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Revision as of 09:53, 18 September 2015

Lab note
Sun. Mon. Tue. Wed. Thu. Fri. Set.
1

PCR
transform 		2

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Sun. Mon. Tue. Wed. Thu. Fri. Set.
1

 2

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5

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 9

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 11
12

 13

 14

 15

Transformation of
GBP[FadL-GBP]
Cultivation of
OmpA[OmpA+pSB1K3] Kr 		16

Cultivation of
GBP[FadL-GBP]
Plasmid purification of
OmpA[OmpA+pSB1K3] Kr
Digestion of
OmpA[OmpA+pSB1K3] Kr (EcoRI and PstI) 		17

Plasmid purification of
GBP[FadL-GBP]
Digestion of
GBP[FadL-GBP] (EcoRI and PstI)
Electrophoresis of
digested product:
GBP[FadL-GBP] (EcoRI and PstI) 		18

Transformation of
ligated product:
GBP[FadL-GBP+pSB1C3] Cr
19

Cultivation of
ligated product: GBP[FadL-GBP] Cr
PCR product: Terminator[J61048] 		20

 21

Cultivation of
GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar
OmpA[OmpA+pSB1C3] Cr
GBP[FadL-GBP+pSB1C3] Cr 		22

Plasmid purification of
GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar
OmpA[OmpA+pSB1C3] Cr
GBP[FadL-GBP+PSB1C3] Cr
Digestion of
GFP[J23101+B0034+E0040+J61048] Ar (EcoRI and PstI)
OmpA[OmpA] Cr (EcoRI and PstI)
GBP[FadL-GBP] Cr (EcoRI and PstI)
Electrophoresis of
digested product:
GFP[J23101+B0034+E0040+J61048] Ar (EcoRI and PstI)
OmpA[OmpA] Cr (EcoRI and PstI)
GBP[FadL-GBP] Cr (EcoRI and PstI) 		23

Digestion of OmpA[OmpA] Cr (EcoRI and SpeI)
GBP[FadL-GBP] Cr (EcoRI and SpeI)
Electrophoresis of
digested product:
OmpA[OmpA] Cr (EcoRI and SpeI)
GBP[FadL-GBP] Cr (EcoRI and SpeI) 		24

Digestion of
OmpA[OmpA] Cr (XbaI and PstI)
GBP[FadL-GBP] Cr (XbaI and PstI) 		25

Electrophoresis of
digested product:
OmpA[OmpA] Cr (XbaI and PstI)
GBP[FadL-GBP] Cr (XbaI and PstI)
Digestion of
RBS[B0034] Ar (XbaI and PstI)
backbone[pSB1A3] Ar (EcoRI and PstI)
Terminator[J61048] Ar (XbaI and PstI)
Electrophoresis of
digested product:
RBS[B0034] Ar (XbaI and PstI)
backbone[pSB1A3] Ar (EcoRI and PstI)
Terminator[J61048] Ar (XbaI and PstI)
Ligation of
OmpA[J23101+B0034+OmpA+pSB1A3] Ar
GBP[GBP+B0034+pSB1K3] Kr
OmpA[OmpA+J61048+pSB1K3] Kr
26

Ligation of
RFP[E1010+B0034+pSB1K3] Kr
Transformation
of ligated product:
OmpA[J23101+B0034+OmpA+pSB1A3] Ar
GBP[FadL-GBP+B0034+pSB1K3] Kr
OmpA[OmpA+J61048+pSB1K3] Kr
Electrophoresis of
digested product:
RFP[E1010+J61048+pSB1K3] Kr
[J23101+B0034] 		27

Digestion of
Terminator[J61048] Ar (XbaI and PstI)
PCR of
ligated product:
OmpA[J23101+B0034+OmpA+pSB1A3] Ar
OmpA[OmpA+J61048+pSB1K3] Kr 		28

Electrophoresis of
RFP[E1010+J61048+pSB1K3] Kr
OmpA[J23101+B0034+OmpA+pSB1A3] Ar Transformation of
ligated product:
GBP[B0034+FadL-GBP+pSB1A3] Ar
OmpA[OmpA+J61048+pSB1K3] Kr PCR of
ligated product
OmpA[J23101+B0034+OmpA] Ar 		29

Cultivation of
RFP[E1010+J61048+pSB1K3] Kr PCR of
OmpA[J23101+B0034+OmpA] Ar
Electrophoresis of
OmpA[J23101+B0034+OmpA] Ar Ligation of
GBP[FadL-GBP+J61048+pSB1K3] Kr 		30

Plasmid purification of
RFP[E1010+J61048+pSB1K3] Kr Ligation of
OmpA[J23101+B0034+OmpA] Ar PCR of
OmpA[J23101+B0034+OmpA] Ar Transformation of
mini: Terminator[J61048]
ligated product: OmpA[J23101+B0034+OmpA] Ar
Sun. Mon. Tue. Wed. Thu. Fri. Set.
1

PCR of
Transformation product:
mini: Terminator[J61048]
ligated product: OmpA[J23101+B0034+OmpA] Ar
GBP[B0034+FadL-GBP+pSB1A3] Ar
Electrophoresis of
mini: Terminator[J61048]
ligated product: OmpA[J23101+B0034+OmpA] Ar 		2
3

Ligation of
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar 		4

Ligation of
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
5/4 Transformation off ligated products:
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
Digestion of
RFP[E1010+J61048] (Kr) (XbaI and PstI)
PCR of ligated products:
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
Electrophoresis of PCR products:
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar 		5

Electrophoresis of
digested products:
RFP[E1010+J61048] (Kr) (XbaI and PstI)
OmpA[J23101+B0034+OmpA] Ar (EcoRI and PstI)
GBP[FadL-GBP+J61048] Kr (EcoRI and PstI)
PCR of
OmpA[J23101+B0034+OmpA+pSB1A3] Ar
Cultivation of PCR product:
RFP[E1010+J61048+pSB1K3] Kr
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar 		6

Plasmid purification of
RFP[E1010+J61048+pSB1K3] Kr
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
Digestion of
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)
Cultivation of PCR product:
OmpA[J23101+B0034+OmpA+pSB1A3] Ar
PCR of backbone:
[pSB1C3] Cr
[pSB1A3] Ar
[pSB1K3] Kr
Electrophoresis of
PCR products: OmpA[J23101+B0034+OmpA+pSB1A3] Ar
digested products: GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI) 		7

Digestion of
RFP[E1010+J61048+pSB1K3] Kr (EcoRI and PstI)
RFP[E1010+J61048+pSB1K3] Kr (XbaI and PstI)
[pSB1C3] Cr (EcoRI and PstI)
[pSB1A3] Ar (EcoRI and PstI)
[pSB1K3] Kr (EcoRI and PstI)
OmpA[J23101+B0034+OmpA+pSB1A3] Ar (EcoRI and PstI)
Plasmid purification of
OmpA[J23101+B0034+OmpA+pSB1A3] Ar
[pSB1K3] Kr
Electrophoresis of digested products:
RFP[E1010+J61048+pSB1K3] Kr (EcoRI and PstI)

RFP[E1010+J61048+pSB1K3] Kr (XbaI and PstI)
[pSB1C3] Cr (EcoRI and PstI)
[pSB1A3] Ar (EcoRI and PstI)
[pSB1K3] Kr (EcoRI and PstI)
Ligation of
RFP[B0034+E1010+J61048+pSB1C3] Cr 		8

Electrophoresis of digested products:
OmpA[J23101+B0034+OmpA] Ar (EcoRI and PstI)
Transformation off ligated products:
RFP[B0034+E1010+J61048+pSB1C3] Cr
PCR of
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
Electrophoresis of PCR products:
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
Transformation off
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
[B0030] Ar
[J23110] Ar 		9

PCR of
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
[B0030] Ar
[J23110] Ar
Electrophoresis of PCR products:
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
[B0030] Ar
[J23110] Ar
Cultivation of
[B0030] Ar
[J23110] Ar
Ligation of
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
10

Electrophoresis of
[B0030] Ar
Cultivation of
[B0030] Ar

Plasmid purification of
[J23110] Ar
[B0030] Ar
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar
Digestion of
[B0030] Ar (EcoRI and SpeI)
[J23110] Ar (EcoRI and SpeI)
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)
GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI)
Electrophoresis of digested products:
[B0030] Ar (EcoRI and SpeI)
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)
GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI) 		11

Electrophoresis of digested products:
[J23110] Ar (EcoRI and SpeI)
Ligation of
RFP[B0030+E1010+J61048+pSB1C3] Cr
GFP[B0030+E0040+J61048+pSB1K3] Kr
Transformation off
RFP[B0030+E1010+J61048+pSB1C3] Cr
GFP[B0030+E0040+J61048+pSB1K3] Kr
PCR of
RFP[B0030+E1010+J61048+pSB1C3] Cr
GFP[B0030+E0040+J61048+pSB1K3] Kr 		12

PCR of ligated products:
RFP[B0030+E1010+J61048+pSB1C3] Cr
Cultivation of PCR products:
RFP[B0030+E1010+J61048+pSB1C3] Cr
Plasmid purification of
RFP[B0030+E1010+J61048+pSB1C3] Cr
Digestion of
RFP[B0030+E1010+J61048] Cr (EcoRI and SpeI)
RFP[B0030+E1010+J61048] Cr (EcoRI and PstI)
Electrophoresis of
digested products:
RFP[B0030+E1010+J61048] Cr (EcoRI and SpeI)
RFP[B0030+E1010+J61048] Cr (EcoRI and PstI)
PCR products: RFP[B0030+E1010+J61048+pSB1C3] Cr 		13

Ligation of
[J23110+B0034+pSB1K3] Kr 		14

Cultivation of
OmpA[J23101+B0034+OmpA+pSB1A3] Ar
Transformation off
[J23110+B0034+pSB1K3] Kr
Electrophoresis of digested products:
GBP[J23101+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)
GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI)
[pSB1K3] Kr (EcoRI and PstI) 		15

Digestion of
GBP[B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)
Ligation of
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr 		16

Transformation off
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr
Electrophoresis of
GBP[B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)
GBP[B0034+FadL-GBP+pSB1A3] Ar (XbaI and PstI)
Ligation of
GBP[B0034+FadL-GBP+pSB1C3] Cr
Transformation off
[J23110+B0034+pSB1K3] Kr
17

Ligation of
[J23110+B0034+pSB1K3] Kr
GBP[J23110+B0034+FadL-GBP+pSB1K3] Kr
PCR of
GBP[B0034+FadL-GBP+pSB1C3] Cr
GBP[J23110+B0034+FadL-GBP+pSB1K3] Kr
Electrophoresis of PCR products:
GBP[B0034+FadL-GBP+pSB1C3] Cr
GBP[J23110+B0034+FadL-GBP+pSB1K3] Kr 		18

Cultivation of
GBP[B0034+FadL-GBP+pSB1C3] Cr
Plasmid purification of
GBP[B0034+FadL-GBP+pSB1C3] Cr
Digestion of
GBP[B0034+FadL-GBP] Cr (XbaI and PstI)
RFP[B0030+E1010+J61048] Cr (XbaI and PstI)
GFP[B0030+E0040+J61048] Transformation off
[J23110+B0034+pSB1K3] Kr
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr
GFP[B0030+E0040+J61048] 		19

Ligation of
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr
PCR of
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr
Electrophoresis of
digested products:
RFP[B0030+E1010+J61048] Cr (XbaI and PstI)
GBP[B0034+FadL-GBP] Cr (XbaI and PstI)
PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr
Transformation off
scFv-B[scFv-B+pSB1K3] Kr
scFv-C[scFv-C+pSB1K3] Kr
GFP[B0030+E0040+J61048]
Cultivation of
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr 		20

Cultivation of
OmpA[J23101+B0034+OmpA] Ar
scFv-B[scFv-B+pSB1K3] Kr
scFv-C[scFv-C+pSB1K3] Kr
Plasmid purification of
OmpA[J23101+B0034+OmpA] Ar
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr
scFv-B[scFv-B+pSB1K3] Kr
scFv-C[scFv-C+pSB1K3] Kr
Digestion of
OmpA[J23101+B0034+OmpA] Ar (EcoRI and Pst1)
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr (EcoRI and PstI)
OmpA[J23101+B0034+OmpA] Ar (NcoI and PstI)
scFv-B[scFv-B] Ar (EcoRI and PstI)
scFv-C[scFv-C] Ar (EcoRI and PstI)
Ligation of
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr Transformation off
[J23110+B0034+pSB1K3] Kr
[pSB3C5] 抗性
Electrophoresis of
OmpA[J23101+B0034+OmpA] Ar (EcoRI and PstI) 		21

Digestion of
scFv-B[scFv-B] Ar (EcoRI and PstI)
scFv-C[scFv-C] Ar (EcoRI and PstI)
Electrophoresis of
OmpA[J23101+B0034+OmpA] Ar (NcoI and PstI)
scFv-B[scFv-B] Ar (EcoRI and PstI)
scFv-C[scFv-C] Ar (EcoRI and PstI)
Ligation of
scFv-B[scFv-B+pSB1C3] Cr
scFv-C[scFv-C+pSB1C3] Cr
scFv-B[scFv-B+pSB1K3] Kr
scFv-C[scFv-C+pSB1K3] Kr 		22

Cultivation of
[pSB3C5] Cr
[J23110+B0034+pSB1K3] Kr
Transformation off
GBP[J23110+B0034+GBP] Ar
PCR of
scFv-B[scFv-B+pSB1C3] Cr
scFv-C[scFv-C+pSB1C3] Cr
scFv-B[scFv-B+pSB1K3] Kr
scFv-C[scFv-C+pSB1K3] Kr
Electrophoresis of
scFv-B[scFv-B+pSB1C3] Cr
scFv-C[scFv-C+pSB1C3] Cr
scFv-B[scFv-B+pSB1K3] Kr
scFv-C[scFv-C+pSB1K3] Kr
Ligation of
scFv-B[scFv-B+pSB1A3] Ar
scFv-C[scFv-C+pSB1A3] Ar
Transform of
scFv-B[scFv-B+pSB1A3] Ar
scFv-C[scFv-C+pSB1A3] Ar 		23

Plasmid purification of
[pSB3C5] Cr
[J23110+B0034+pSB1K3] Kr
Digestion of
[pSB3C5] Cr (EcoRI and PstI)
[J23110+B0034+pSB1K3] Kr (EcoRI and PstI)
PCR of
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr
Transformation off
scFv-B[scFv-B+pSB1A3] Ar
scFv-C[scFv-C+pSB1A3] Ar
24

Electrophoresis of digested products:
[pSB3C5] Cr (EcoRI and PstI)
[J23110+B0034+pSB1K3] Kr (EcoRI and PstI)
Cultivation of
PCR products: GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr
scFv-B[scFv-B+pSB1C3] Cr
scFv-C[scFv-C+pSB1A3] Ar
Plasmid purification of
scFv-B[scFv-B+pSB1A3] Ar
scFv-C[scFv-C+pSB1A3] Ar
GFP[B0030+E0040+J61048]
Digestion of
[J23101+B0034] Kr (EcoRI and SpeI)
Ligation of
OmpA[J23101+B0034+OmpA+pSB3C5] Cr
PCR of
GBP[J23110+B0034+GBP] Ar by two pArts
Electrophoresis of
GBP[J23110+B0034+GBP] Ar 		25

Transformation off
OmpA[J23101+B0034+OmpA+pSB3C5] Cr
Cultivation of
GBP[J23110+B0034+FadL-GBP] Ar
Digestion of
GBP[B003+FadL-GBP] Cr (XbaI and PstI)
scFv-B[scFv-B+pSB1C3] Cr (EcoRI and PstI)
scFv-C[scFv-C+pSB1A3] Ar (EcoRI and PstI)
GFP[B0030+E0040+J61048] (XbaI and PstI)
Electrophoresis of
GFP[B0030+E0040+J61048]
scFv-B[scFv-B+pSB1C3] Cr (EcoRI and PstI)
scFv-C[scFv-C+pSB1A3] Ar (EcoRI and PstI)
GFP[B0030+E0040+J61048] (XbaI and PstI)
Cultivation of
scFv-B[scFv-B+pSB1C3] Cr (EcoRI and PstI)
scFv-C[scFv-C+pSB1A3] Ar (EcoRI and PstI)
GFP[B0030+E0040+J61048] (XbaI and PstI) 		26

Ligation of
OmpA[J23110+B0034+OmpA+pSB1A3] Ar
GBP[J23110+B0034+GBP+pSB1A3] Ar
Gel extraction of
[J23110] Ar (SpeI and PstI)
Plasmid purification of
GBP[J23110+B0034+GBP] Ar by two pArts
Digestion of
GBP[J23110+B0034+GBP] Ar by two pArts (EcoRI and SpeI) 		27

PCR of
[J23101+B0034+OmpA+pSB3C5]
Electrophoresis of
GBP[J23110+B0034+GBP] Ar by two pArts (EcoRI and SpeI)
GBP[B0034+FadL-GBP] Cr (XbaI and PstI)
[J23110] Ar (SpeI and PstI) 		28

Ligation of
scFv-B[scFv-B+pSB1A3] Ar
scFv-C[scFv-C+pSB1A3] Ar
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048] 抗性??
scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar
Transform of
scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048]
scFv-B[scFv-B+pSB1A3] Ar
scFv-C[scFv-C+pSB1A3] Ar
PCR of
[J23101+B003+OmpA+pSB3C5] + primer(EX-OmpA and OmpA-S)
OmpA[J23101+B0034+OmpA+pSB3C5] Cr

Plasmid purification in gel slices of PCR products:
OmpA[EX-OmpA-S]
Gel extraction of
OmpA[OmpA] (EcoRI and SpeI)
Transformation off
OmpA[J23101+B0034+OmpA+pSB3C5] 		29

Plasmid purification of
[J23110] Ar
[pSB1C3]
Digestion of
[J23110] Ar (EcoRI and SpeI)
[pSB1C3] (EcoRI and SpeI)
Ligation of
scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar
[J23110+B0034+pSB1K3] Kr
Electrophoresis of
[J23110] Ar (EcoRI and SpeI)
[pSB1C3] (EcoRI and SpeI)
Transform of
scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar
[J23110+B0034+pSB1K3] Kr
GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr 		30

PCR of
scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar
[J23110+B0034+pSB1K3] Kr
GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr
31

Cultivation of
[J23110+B0034] Kr
GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar
Plasmid purification of
[J23110+B0034] Kr
GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar
Digestion of
[J23110+B0034] Kr (EcoRI and PstI)
GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)
scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar (EcoRI and PstI)
[J23110+B0034] (EcoRI and SpeI) Kr
[J23101+B0034] (EcoRI and SpeI) Kr
OmpA[OmpA] Cr (EcoRI and PstI)
[pSB1K3] (EcoRI and PstI)
Electrophoresis of
[J23110+B0034] Kr (EcoRI and PstI)
GFP[J23110+B0034+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)
scFv-C[J23101+B0034+OmpA+scFv-C+pSB1A3] Ar (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B+pSB1A3] Ar (EcoRI and PstI)
Ligation of
OmpA[OmpA+pSB1C3] Cr
Sun. Mon. Tue. Wed. Thu. Fri. Set.
1

Electrophoresis of
[J23110+B0034] (EcoRI and SpeI) Kr
[J23101+B0034] (EcoRI and SpeI) Kr
OmpA[OmpA] (NcoI and PstI)
GFP[K1489002+E0040] Kr
Digestion of
[pSB1C3] (EcoRI and PstI)
[pSB1A3] (EcoRI and PstI)
OmpA[OmpA] (EcoRI and NcoI)
[pSB3C5] (EcoRI and PstI)
Ligation of
GFP[J23101+B0034+K1489002+E0040+pSB3C5] (Cr)
GFP[J23110+B0034+K1489002+E0040+pSB3C5] (Cr)
Electrophoresis of digested product:
[pSB1C3] (EcoRI and PstI)
[pSB1A3] (EcoRI and PstI)
OmpA[OmpA] (EcoRI and NcoI)
Cultivation of
BFP[OmpA+K592100]
PCR of
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr
Electrophoresis of
GBP[J23101+B0034+FadL-GBP+pSB1K3] Kr 		2

PCR of
OmpA[EX-OmpA-S+pSB1C3] Cr
Electrophoresis of
OmpA[EX-OmpA-S+pSB1C3] Cr
OmpA[OmpA] (EcoRI and NcoI)
Ligation of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E1010+J61048+pSB1K3] Kr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr
Digestion of
[pSB1A3] (EcoRI and NcoI)
Electrophoresis of digested products:
[pSB1A3] (EcoRI and NcoI) 		3

Ligation of
OmpA[OmpA+psB1C3] Cr
[J23110+B0034+pSB1K3] Kr
Transform of
OmpA[OmpA+psB1C3] Cr
[J23110+B0034+pSB1K3] Kr
Digestion of
BFP[OmpA+ K592100] (NcoI and PstI)
OmpA[OmpA] (NcoI and PstI) 		4

Cultivation of
[J23110+pSB1A3] Ar
Transform of
[pSB1C3]
Digestion of
[pSB1C3] (EcoRI and PstI)
Ligation of
OmpA[OmpA+pSB1C3] Cr
OmpA[J23101+B0034+OmpA+pSB3C5] Cr
Transform of
[B0034+pSB1A3] Ar 		5

Transform of
OmpA[J23101+B0034+OmpA+pSB3C5] Cr
Plasmid purification of
[J23110+pSb1A3] Ar
Digestion of
[J23110+pSb1A3] Ar
PCR of
[J23110+pSb1A3] Ar
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
Transform of
OmpA[OmpA+pSB1C3] Cr
Electrophoresis of
[pSB1C3]
[B0034+pSB1A3] Ar
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr 		6

Cultivation of
[pSB1C3]
Digestion of
[pSB1C3] (EcoRI and SpeI)
7

Gel extraction of
OmpA[EX-OmpA-S] 抗性
[pSB1C3] (EcoRI and SpeI)
Gel extraction of
[pSB1C3] (EcoRI and SpeI)
Transform of
GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar
[pSB1A3]
OmpA[J23101+B0034+OmpA] Ar
Ligation of
OmpA[OmpA+pSB1C3] Cr
Cultivation of
[pSB1C3] 		8

Transform of
OmpA[OmpA+pSB1C3] Cr
[pSB3K3] [
pSB6A1]
Plasmid purification of
[pSB1C3]
Digestion of
OmpA[EX-OmpA-S] (EcoRI and SpeI)
Electrophoresis of
[pSB1C3] (EcoRI and SpeI)
Transform of
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr 		9

Ligation of
OmpA[OmpA+pSB1C3] Cr
PCR of
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr
Cultivation of
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
Plasmid purification of
[pSB3K3]
[pSB6A1]
Digestion of
[pSB3K3] (EcoRI and PstI)
[pSB6A1] (EcoRI and PstI) 		10

Transform of
[OmpA+pSB1C3] Cr
Plasmid purification of
[J23101+B0034+OmpA+K592100+pSB3C5] Cr
Digestion of
[J23101+B0034+OmpA+K592100] Cr (EcoRI and PstI)
Electrophoresis of
[pSB3K3] (EcoRI and PstI)
[pSB6A1] (EcoRI and PstI)
[pSB1A3] (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B] (EcoRI and PstI)
scFv-C[J23101+B0034+OmpA+scFv-C] (EcoRI and PstI)
Transform of
[J23110+B0034] Kr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3]
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3]
Ligation of
scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr
Electrophoresis of
[pSB3K3]
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr 		11

Transform of
scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr
[J23110+B0034+pSB1C3] Cr
Ligation of
[J23110+B0034+pSB1K3] Kr
[J23110+B0034+pSB1C3] Cr
PCR of
OmpA[OmpA+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr
Electrophoresis of
OmpA[OmpA+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr
Cultivation of
OmpA[OmpA+pSB1C3] Cr 		12

PCR of
[J23110+B0034+pSB1C3] Cr
[B0034+pSB1A3] Ar
Plasmid purification of
OmpA[OmpA+pSB1C3] Cr
Ligation of
[J23110+B0034+pSB1C3] Cr
Transform of
[J23110+B0034+pSB1C3] Cr 		13

Electrophoresis of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)
14

SDS-page of
[pSB1C3]
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr (EcoRI and PstI)
[pSB3C5]
scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr
scFv-B[OmpA+scFv-B]
scFv-C[OmpA+scFv-C]
Electrophoresis of
[J23110]
[B0034+FadL-GBP+B0030+E1010+J61048] 抗性 (XbaI and PstI)
Ligation of
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB3K3] Cr
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Cr
Digestion of
OmpA[OmpA+pSB1C3] Cr (EcoRI and SpeI)
OmpA[OmpA+pSB1C3] Cr (EcoRI and PstI)
Transform of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
Ligation of
GBP[FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar
Transform of
GBP[FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar 		15

 16

Transform of
[pSB3C5] 		17

Cultivation of
[pSB3C5] 		18

Plasmid purification of
[pSB3C5]
Digestion of
[pSB3C5]( EcoRI and PstI)
Electrophoresis of
[pSB3C5]( EcoRI and PstI)
Ligation of
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr 		19

Transform of
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr 		20

PCR of
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr
Electrophoresis of
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr
21

 22

 23

PCR of
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr
Cultivation of
[pSB3C5]
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
BFP[OmpA+K592100]
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1]
[psB6A1]
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar
[pSB1A3]
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1C3] Cr
Ligation of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar 		24

Plasmid purification of
[pSB3C5] 		25

Cultivation of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
Transform of
[J23101+B0034+pSB1K3] Kr 		26

 27

Digestion of
OmpA[OmpA] Cr (EcoRI and PstI)
Electrophoresis of
GBP[FadL-GBP] 抗性 (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ar (EcoRI and PstI)
OmpA[OmpA] Cr (EcoRI and PstI)
28

Cultivation of
BFP[J23101+B0034+OmpA+K592100+pSB1A3] Ar
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB3C5] Cr
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
[pSB6A1]
[pSB3C5]
[pSB1C3]
[pSB1A3]
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
Electrophoresis of
[J23101+B0034] Kr
GBP[FadL-GBP] 抗性 (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ar (EcoRI and PstI)
OmpA[OmpA] Cr (EcoRI and PstI)
BFP[J23101+B0034+OmpA+K592100] Ar (EcoRI and PstI)
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Cr (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Cr (EcoRI and PstI)
[pSB6A1] (EcoRI and PstI)
OmpA[OmpA] Cr (XbaI and PstI)
[E0040] Ar (XbaI and PstI)
Ligation of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
[B0034+E0040+pSB1K3] Kr
GBP[FadL-GBP+pSB6A1] Ar
GBP[FadL-GBP+pSB1C3] Cr
GBP[FadL-GBP+pSB3C5] Cr
Digestion of
BFP[J23101+B0034+OmpA+K592100] Cr (XbaI and PstI)
SDS-page of
[pSB1A3]
[pSB1C3]
BFP[J23101+B0034+OmpA+K592100+pSB1A3] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
[pSB3C5]
[pSB6A1]
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB3C5] Cr
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar
Transform of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
GBP[FadL-GBP+pSB1C3] Cr
GBP[FadL-GBP+pSB3C5] Cr
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr 		29

PCR of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
GBP[FadL-GBP+pSB1C3] Cr
GBP[FadL-GBP+pSB3C5] Cr
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
Electrophoresis of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar 		30

Electrophoresis of
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
Sun. Mon. Tue. Wed. Thu. Fri. Set.
1

Cultivation of
[pSB6A1]
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
Transform of
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
Digestion of
GBP[]FadL-GBP] 抗性 (EcoRI and PstI) 		2

Digestion of
[J23110] Ar (SpeI and PstI)

Plasmid purification in gel slices of
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
Digestion of
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)
scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)
Electrophoresis of
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)
scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) 		3

Gel extraction of
[J23110] Ar (SpeI and PstI)
[pSB6A1] (EcoRI and PstI)
Transform of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
Electrophoresis of
GBP[FadL-GBP] (EcoRI and Pstl)
Ligation of
GBP[FadL-GBP+pSB3C5] Cr
GBP[FadL-GBP+pSB1C3] Cr
GBP[FadL-GBP+pSB6A1] Ar 		4

Transform of
GBP[FadL-GBP+pSB3C5] Cr
GBP[FadL-GBP+pSB1C3] Cr
GBP[FadL-GBP+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
Electrophoresis of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
Cultivation of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
[J23101+B0034+pSB1K3] Kr
5

Plasmid purification of
[J23101+B0034+pSB1K3] Kr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr 		6

Cultivation of
[pSB1C3]
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
[pSB6A1]
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
[pSB1A3]
GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar 		7

Digestion of
[J23101+B0034+pSB1K3] Kr (EcoRI and PstI)
SDS-page of
[pSB1C3]
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
[pSB6A1]
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
[pSB1A3]
GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar
Ligation of
[J23110+B0034] Ar
Electrophoresis of
[J23101+B0034+pSB1K3] Kr (EcoRI and PstI)
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar 		8

Plasmid purification of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
Transform of
[J23110+B0034] Ar
BFP[J23101+B0034+OmpA+K592100]
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100]
Electrophoresis of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) 		9

Electrophoresis of
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+scFv-B+B0034+E0040+J61048+pSB1C3] Cr
[pSB3C5]
BFP[J23101+B0034+OmpA+scFv-B+B0030+e0040+J61048+pSB6A1] Ar
Cultivation of
scFv-B[J23110+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar 		10

Plasmid purification of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar 		11

PCR of
[J23110+B0034] Ar
Electrophoresis of
[J23110+B0034] Ar
12

Cultivation of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
[J23110+B0034] Ar
Plasmid purification of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
Digestion of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) 		13

Plasmid purification of
[J23110+B0034] Ar
Transform of
[R0010]
Electrophoresis of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) 		14

Digestion of
[J23110+B0034] Ar (EcoRI and PstI)
GBP[FadL-GBP] Kr (XbaI and PstI)
OmpA[OmpA] Cr (EcoRI and SpeI)

Electrophoresis of
[J23110+B0034] Ar (EcoRI and PstI)
GBP[FadL-GBP] Kr (XbaI and PstI)
OmpA[OmpA] Cr (EcoRI and SpeI)
Transform of
[R0010] Cr
Cultivation of
[pSB1C3]
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr 		15

Digestion of
[pSB6A1]
[pSB3C5]
Anti-Her 2[Herceptin+E1010]
[J23101+B0034+OmpA+scFv-C]
[J23101+B0034+OmpA+scFv-B]
Cultivation of
[pSB6A1]
[R0010]
PCR of
OmpA[OmpA-linker] (EcoRI and PstI)
Transform of
[J23110+B0034] Kr
BFP[K592100+J61048] Kr 		16

Gel extraction of
OmpA[OmpA-linker] (EcoRI and PstI)
Ligation of
OmpA[J23101+B0034+OmpA+pSB3C5] Cr
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
Digestion of
OmpA[OmpA-linker] (EcoRI and PstI)
Transform of
OmpA[J23101+B0034+OmpA+pSB3C5] Cr 		17

Electrophoresis of
OmpA[OmpA-linker] (EcoRI and PstI)
Anti-Her 2[Herceptin+E1010] (EcoRI and PstI)
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI) 		18

Ligation of
OmpA[OmpA-linker+pSB1C3] Cr
Anti-Her 2[Herceptin+E1010+pSB1C3] Cr
19

Transform of
OmpA[OmpA-linker+pSB1C3] Cr
Anti-Her 2[Herceptin+E1010+pSB1C3] Cr 		20

 21

 22

 23

 24

Transform of
OmpA[OmpA-linker+pSB1C3] Cr
Anti-Her 2[Herceptin+E1010+pSB1C3] Cr
PCR of
OmpA[OmpA-linker+pSB1C3] Cr
Anti-Her 2[Herceptin+E1010+pSB1C3] Cr
Electrophoresis of
OmpA[OmpA-linker+pSB1C3] Cr
Anti-Her 2[Herceptin+E1010+pSB1C3] Cr 		25

Cultivation of
[R0010] Cr
BFP[K592100+J61048] Kr
PCR of
OmpA[OmpA-linker+pSB1C3] Cr
Anti-Her 2[Herceptin+E1010+pSB1C3] Cr
Electrophoresis of
OmpA[OmpA-linker+pSB1C3] Cr
Anti-Her 2[Herceptin+E1010+pSB1C3] Cr
26

Plasmid purification of
[R0010] Cr
BFP[K592100+J61048] Kr
Digestion of
[R0010] Cr (EcoRI and SpeI)
BFP[K592100+J61048] Kr (EcoRI and PstI)
Electrophoresis of
[R0010] Cr
BFP[K592100+J61048] Kr
Ligation of
[R0010+B0034+pSB1K3] Kr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr
BFP[B0030+K592100+J61048+pSB1C3] Cr
Transform of
scFv-C[J23101+B0034+OmpA+scFv-C] Ar
Ligation of
OmpA[OmpA-linker+pSB1C3] Cr
PCR of
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048]
Cultivation of
[pSB1C3]
GFP[J23101+B0034+E0040+J61048+pSB1A3] Ar scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr
GBP[FadL-GBP+pSB6A1] Ar 		27

PCR of
GBP[FadL-GBP]
Anti-Her 2[Herceptin+E1010]
Gel extraction of
GBP[FadL-GBP]
Anti-Her 2[Herceptin+E1010]
Transform of
[R0010+B0034+pSB1K3]
GBP[R0010+B0034+FadL-GBP+B0030+J61048+pSB1K3]
BFP[B0030+K592100+J61048+pSB1C3] Cr
PCR of
GBP[J23110+B0034+FadL-GBP+E0040+J61048] Cr
Gel extraction of
GBP[J23110+B0034+FadL-GBP+E0040+J61048] Cr
SDS-page of
[pSB1C3]
GFP[J23101+B0034+E0040+J61048+pSB1C3] Cr scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr
GBP[J23110+B0034+FadL-GBP+E0040+J61048+pSB1C3] Cr
GBP[FadL-GBP+pSB6A1] Ar
[pSB6A1] 		28

Ligation of
[R0010+B0034+pSB1K3]
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB1K3] Kr
BFP[B0030+K592100+J61048+pSB1C3] Cr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB1C3] Cr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB6A1] Ar
PCR of
BFP[B0030+K592100+J61048+pSB1C3] Cr
[R0010+B0034+pSB1K3]
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB1K3] Kr
OmpA[J23110+B0034+OmpA+pSB3C5] Cr
Digestion of
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048] (XbaI and PstI)
Electrophoresis of
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048] (XbaI and PstI)
SDS-page of
[pSB1C3]
BFP[J23101+B0034+OmpA+K592100+pSB1C3] Cr
GFP[J23101+B0034+E0040+J61048+pSB1C3] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1C3] Cr
[pSB6A1]
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB6A1] Ar
Transform of
OmpA[OmpA-linker] Cr 		29

Electrophoresis of
OmpA[J23101+B0034+OmpA+pSB3C5]
Transform of
GBP[J23110+B0034+FadL-GBP+B0030+E0040+J61048+pSB1C3] Cr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J661048+pSB6A1] Ar
PCR of
BFP[B0034+K592100+J61048+pSB1C3] Cr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]
Electrophoresis of
BFP[B0034+K592100+J61048+pSB1C3] Cr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]
Digestion of
Anti-Her 2[Herceptin-own] (EcoRI and PstI)
Anti-Her 2[Herceptin-E1010] (EcoRI and PstI)
GBP[GBP] (EcoRI and PstI)
Ligation of
[R0010+B0034+pSB1K3] Kr
BFP[B0030+K592100+J61048] Cr
GBP[R0010+B0034+FadL-GBP+B0034+E0040+J61048] Ar
GFP[R0010+B0030+E0040+J61048] Ar
Electrophoresis of
Anti-Her 2[Herceptin-own] (EcoRI and PstI)
Anti-Her 2[Herceptin-E1010] (EcoRI and PstI)
GBP[GBP] (EcoRI and PstI)
[R0010+B0034+pSB1K3] Kr
BFP[B0030+K592100+J61048] Cr
GBP[R0010+B0034+FadL-GBP+B0034+E0040+J61048] Ar
GFP[R0010+B0030+E0040+J61048] Ar 		30

Cultivation of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB1C3] Cr
Ligation of
GBP[FadL-GBP+pSB1C3]Cr
Digestion of
Anti-Her 2[Herceptin own] (NcoI and PstI)
[pSB1A3] (EcoRI and PstI)
[pSB1C3] (EcoRI and PstI)
Cultivation of
scFv-C[J23101+B0034+OmpA+scFv-C] Ar
[J23101+B0034] Kr
Digestion of
scFv-B[scFv-B] Kr (NcoI and PstI)
scFv-C[scFv-C] Kr (NcoI and PstI)
OmpA[OmpA] Cr (EcoRI and PstI)
Electrophoresis of
Anti-Her 2[Herceptin own] (NcoI and PstI)
[pSB1A3] (EcoRI and PstI)
[pSB1C3] (EcoRI and PstI)
Plasmid purification of
scFv-C[J23101+B0034+OmpA+scFv-C] Ar
[J231-1+B0034] Kr
Digestion of
scFv-C[J23101+B0034+OmpA+scFv-C] Ar (EcoRI and SpeI)
[J231-1+B0034] Kr (EcoRI and SpeI)
Anti-Her 2[Herceptin-Own] (NcoI and PstI) 		31

Electrophoresis of
scFv-C[J23101+B0034+OmpA+scFv-C] Ar (EcoRI and SpeI)
[J231-1+B0034] Kr (EcoRI and SpeI)
Anti-Her 2[Herceptin-Own] (NcoI and PstI)
Ligation of
Anti-Her 2[J2310+B0034+OmpA+Herceptin-Own+pSB1A3] Ar
Transform of
Anti-Her 2[J2310+B0034+OmpA+Herceptin-Own+pSB1A3] Ar
Electrophoresis of
[pSB1C3] (XbaI and PstI)
OmpA[OmpA] Cr (EcoRI and PstI)
scFv-B[scFv-B] Kr (NcoI and PstI)
scFv-C[scFv-C] Kr (NcoI and PstI)
Sun. Mon. Tue. Wed. Thu. Fri. Set.
1

Digestion of
Anti-Her 2[Herceptin-Web] (NcoI and PstI)
Electrophoresis of
Anti-Her 2[Herceptin-Web] (NcoI and PstI)
2

Electrophoresis of
[pSB1C3] (XbaI and PstI)
OmpA[OmpA] (EcoRI and PstI)
scFv-B[scFv-B] Kr (NcoI and PstI)
scFv-C[scFv-C] Kr (NcoI and PstI)
OmpA[OmpA] (NcoI and PstI)
Digestion of
BFP[OmpA+K592100] Cr (EcoRI and NcoI) 		3

Transform of [R0010+B0034+pSB1K3] Kr
GBP[FadL-GBP+pSB1C3] Cr
BFP[B0030+K592100+J61048] Cr
Digestion of
Anti-Her 2[Herceptin-Web] (NcoI and PstI)
Ligation of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar 		4

Digestion of
BFP[OmpA+K592100] Cr (EcoRI and NcoI)
PCR of
[R0010+B0034+pSB1K3] Kr
BFP[B0030+K592100+J61048] Cr
Transform of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar 		5

Digestion of
[pSB6A1] (EcoRI and PstI)
[pSB1C3] (EcoRI and PstI)
PCR of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar
Electrophoresis of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar
Cultivation of
BFP[B0030+K592100+J61048] Cr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]
PCR of
GBP[FadL-GBP] Cr
BFP[OmpA+K592100] Cr
scFv-C[J23101+B0034+OmpA+scFv-C] Ar
scFv-C[scFv-C+pSB1C3] Cr
scFv-B[scFv-B+oSB1C3] Cr
Digestion of
scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr (EcoRI and PstI)
scFv-B[J23101+B0034+OmpA+scFv-B+pSB3C5] Cr (EcoRI and PstI)
Transform of
GBP[J23101+B0034+FadL-GBP]Cr
scFv-B[scFv-B+pSB1C3] Cr
scFv-C[scFv-C+pSB1C3] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Cr
Gel extraction of
[pSB6A1] (EcoRI and PstI)
[OmpA] (NcoI and PstI)
[pSB1C3] (EcoRI and PstI) 		6

Cultivation of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar
Ligation of
Anti-Her 2[OmpA+Herceptin-Web] Cr
Anti-Her 2[OmpA+Herceptin-Own] Cr
scFv-C[OmpA+scFv-C] Cr
scFv-B[OmpA+scFv-B] Cr
scFv-C[J23101+B0034+OmpA+scFv-C] Cr
scFv-B[J23101+B0034+OmpA+scFv-B] Cr
Transform of
Anti-Her 2[OmpA+Herceptin-Web] Cr
Anti-Her 2[OmpA+Herceptin-Own] Cr
scFv-C[OmpA+scFv-C] Cr
scFv-B[OmpA+scFv-B] Cr
scFv-C[J23101+B0034+OmpA+scFv-C] Cr
scFv-B[J23101+B0034+OmpA+scFv-B] Cr
[pSB1C3]
[pSB1A3]
[pSB1K3]
Plasmid purification of
BFP[B0030+K592100+J61048]
Digestion of
BFP[B0030+K592100+J61048] (EcoRI and PstI)
Cultivation of
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Cr
Ligation of
[R0010+B0034] Kr
GBP[GBP[J23101+B0034+FadL-GBP]+pSB1C3] Cr
Digestion of
[pSB1C3] (XbaI and PstI)
Electrophoresis of
GBP[J23101+B0034+FadL-GBP] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040] Cr
scFv-C[scFv-C] Cr
scFv-B[scFv-B] Cr
Plasmid purification of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar
Digestion of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and PstI)
Ligation of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar
Electrophoresis of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar
[B0034]
[pSB1C3]
Cultivation of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar
Transform of
GBP[J23101+B0034+FadL-GBP] Ar
Electrophoresis of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040] Cr
[pSB1C3] (XbaI and PstI)
[pSB6A1] (XbaI and PstI)
[pSB1C3] (XbaI and PstI)
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar
Cultivation of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Cr
GFP[J23101+B0034+E0040+J61048] Ar
SDS-page of
GFP[J23101+B0034+E0040+J61048] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Ar
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048] Ar
[pSB6A1]
GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Ar
Digestion of
[pSB1C3] (XbaI and PstI)
Anti-Her 2[Herceptin-Web] (NcoI and PstI)
Anti-Her 2[Herceptin-Own] (NcoI and PstI)
Ligation of
[pSB1C3] (XbaI and PstI)
Anti-Her 2[Herceptin-Web] Cr
Anti-Her 2[Herceptin-Own] Cr
BFP[J23101+B0034+OmpA+K592100] Cr 		7

Plasmid purification of
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Cr
Digestion of
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Cr
PCR of
scFv-B[J23101+B0034+OmpA+scFv-B] Cr
scFv-C[J23101+B0034+OmpA+scFv-C] Cr
Electrophoresis of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and PstI)
Cultivation of
scFv-B[J23101+B0034+OmpA+scFv-B] Cr
scFv-C[J23101+B0034+OmpA+scFv-C] Cr
Transform of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar
Plasmid purification of
GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Cr
Digestion of
GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Cr (EcoRI and PstI)
PCR of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar
Electrophoresis of
scFv-B[J23101+B0034+OmpA+scFv-B] Cr
scFv-C[J23101+B0034+OmpA+scFv-C] Cr
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar
OmpA[OmpA] (NcoI and PstI)
[pSB1C3] (EcoRI and PstI)
Ligation of
scFv-C[OmpA+scFv-C] Cr
scFv-B[OmpA+scFv-B] Cr
Anti-Her 2[Herceptin-Own] Cr
Anti-Her 2[Herceptin-Web] Cr
Anti-Her 2[OmpA+Herceptin-Own] Cr
Anti-Her 2[OmpA+Herceptin-Web] C
Electrophoresis of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and PstI)
Anti-Her 2[Herceptin-Web] (NcoI and PstI)
Anti-Her 2[Herceptin-Own] (NcoI and PstI)
Transform of
scFv-C[OmpA+scFv-C] Cr
scFv-B[OmpA+scFv-B] Cr
Anti-Her 2[Herceptin-Own] Cr
Anti-Her 2[Herceptin-Web] Cr
Anti-Her 2[OmpA+Herceptin-Own] Cr
Anti-Her 2[OmpA+Herceptin-Web] C
BFP[J23101+B0034+OmpA+K592100] Cr 		8

Plasmid purification of
scFv-C[J23101+OmpA+scFv-C] Cr
scFv-B[J23101+OmpA+scFv-B] Cr
Digestion of
scFv-C[J23101+OmpA+scFv-C] Cr (EcoRI and PstI)
scFv-B[J23101+OmpA+scFv-B] Cr (EcoRI and PstI)
PCR of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar
Cultivation of
[pSB1K3]
[pSB1C3]
[pSB1A3]
PCR of
BFP[J23101+B0034+OmpA+K592100] Cr
Transform of
scFv-C[OmpA+scFv-C] Cr
Electrophoresis of
BFP[J23101+B0034+OmpA+K592100] Cr
scFv-C[J23101+OmpA+scFv-C] Cr (EcoRI and PstI)
scFv-B[J23101+OmpA+scFv-B] Cr (EcoRI and PstI)
[pSB1C3] (EcoRI and PstI)
Digestion of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar (EcoRI and SpeI)
OmpA[OmpA] Cr NcoI and PstI
OmpA[OmpA+K592100] Cr NcoI and PstI
Ligation of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Cr
Digestion of
Anti-Her 2[Herceptin-Own] (EcoRI and PstI)
Anti-Her 2[Herceptin-Web] (EcoRI and PstI)
Electrophoresis of
BFP[J23101+B0034+OmpA+K592100] Cr
9

Cultivation of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar
Plasmid purification of
[pSB1C3]
[pSB1K3]
[pSB1A3]
Digestion of
[pSB1C3] (EcoRI and PstI)
[pSB1K3] (EcoRI and PstI)
[pSB1A3] (EcoRI and PstI)
Transform of
Anti-Her 2[Herceptin-Own] Cr
Anti-Her 2[Herceptin-Web] Cr
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Cr
[R0010+B0034] Cr
BFP[J23101+B0034+OmpA+K592100] Cr
Electrophoresis of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Cr (EcoRI and SpeI)
OmpA[OmpA] Cr (NcoI and PstI)
Digestion of
[pSB1A3] (EcoRI and PstI)
[pSB1K3] (EcoRI and PstI)
[pSB1C3] (EcoRI and PstI)
RFP[B0030+E1010+J61048] Cr (XbaI and PstI)
Electrophoresis of
scFv-C[OmpA+scFv-C] Kr
PCR of
GBP[GBp+pSB1C3] Cr
Digestion of
BFP[B0030+K592100+J61048] Cr (XbaI and PstI) 		10

Electrophoresis of
GBP[GBp+pSB1C3] Cr
[pSB1A3] (EcoRI and PstI)
[pSB1K3] (EcoRI and PstI)
[pSB1C3] (EcoRI and PstI)
Ligation of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web+B0030+E1010+J61048] Kr
Cultivation of
[R0010+B0034] Cr
BFP[B0030+K592100+J61048] Cr
scFv-C[OmpA+scFv-C] Kr
Digestion of
[pSB1C3] (EcoRI and Pstl)
Transform of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048+pSB1K3] Kr
Gel extraction of
[pSB1C3] (EcoRI and PstI)
PCR of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web]
Anti-Her 2[Herceptin-Web]
Anti-Her 2[Herceptin-Own] 		11

Plasmid purification of
[R0010+B0034] Cr
scFv-C[OmpA+scFv-C] Kr
BFP[OmpA+K592100] Cr

Gel extraction of
[pSB1C3] (EcoRI and PstI)
Digestion of
[R0010+B0034] Cr (EcoRI and SpeI)
scFv-C[OmpA+scFv-C] Kr (EcoRI and PstI)
BFP[OmpA+K592100] Cr (EcoRI and PstI)
GBP[J23101+B0034+FadL-GBP] Cr (EcoRI and SpeI)
Ligation of
Anti-Her 2[Herceptin-Web] Cr
PCR of
scFv-B[OmpA+scFv-B] Kr
Electrophoresis of
[R0010+B0034] Cr (EcoRI and SpeI)
scFv-C[OmpA+scFv-C] Kr (EcoRI and PstI)
BFP[OmpA+K592100] Cr (EcoRI and PstI)
GBP[J23101+B0034+FadL-GBP] Cr (EcoRI and SpeI)
Ligation of
GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar
Electrophoresis of
scFv-B[OmpA+scFv-B] Kr
PCR of
scFv-C[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr
Transform of
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr 		12

Electrophoresis of
scFv-B[OmpA+scFv-B] Kr
Transform of
Anti-Her 2[Herceptin-Web] Cr
Anti-Her 2[Herceptin-Own] Cr
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Web] Ar
PCR of
scFv-C[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
Digestion of
[pSB1C3] (EcoRI and PstI)
Electrophoresis of
scFv-C[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar 		13

Transform of
GBP[J231010+B0034+FadL-GBP+B0030+E0040+J61048+pSB6A1] Ar
Cultivation of
[pSB1C3]
scFv-B[OmpA-scFv-B] Kr
Plasmid purification of
[pSB1C3]
scFv-B[OmpA-scFv-B] Kr
Digestion of
[pSB1C3] (EcoRI and PstI)
scFv-B[OmpA-scFv-B] Kr (EcoRI and PstI)
Electrophoresis of
[pSB1C3] (EcoRI and PstI)
Ligation of
OmpA[R0010+B0034+OmpA+pSB1A3] Ar
GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar
scFv-B[J23101+B0034+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
Electrophoresis of
[pSB1C3] (EcoRI and PstI)
scFv-B[OmpA-scFv-B] Kr (EcoRI and PstI) 		14

Transform of
OmpA[R0010+B0034+OmpA+pSB1A3] Ar
GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar 		15

PCR of
OmpA[R0010+B0034+OmpA+pSB1A3] Ar
GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar
scFv-C[J23101+B0034+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
Ligation of
scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr
BFP[J23101+B0034+OmpA+K592100+pSB3C5] Cr
PCR of
GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr
Transform of
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr
Electrophoresis of
GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr
Electrophoresis of
GBP[FadL-GBP] (EcoRI and PstI)
16

PCR of
GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Kr
Cultivation of
OmpA[R0010+B0034+OmpA]
Electrophoresis of
GBP[R0010+B0034+FadL-GBP+pSB1A3] Kr
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048] Kr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048] Kr 		17

Ligation of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar
Transform of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048+pSB6A1] Ar 		18

Plasmid purification of
OmpA[R0010+B0034+OmpA+pSB1A3] Ar
GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar
Digestion of
BFP[OmpA+K592100] Cr (EcoRI and NcoI)
Anti-Her 2[Herceptin-Own] (NcoI and SpeI)
Anti-Her 2[Herceptin-Web] (NcoI and SpeI)
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar (EcoRI and SpeI)
BFP[B0030+K592100+J61048] Cr (XbaI and PstI)
OmpA[R0010+B0034+OmpA+pSB1A3] Ar (EcoRI and PstI)
GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar (EcoRI and PstI)
Ligation of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own+pSB1C3] Cr
GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar
Transform of
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own+pSB1C3] Cr
GBP[R0010+B0034+FadL-GBP+pSB1A3] Ar
GBP[J23101+B0034+fadL-GBP+B0030+E0040+J61048] Cr
OmpA[R0010+B0034+OmpA+pSB1C3] Cr
GBP[R0010+B0034+FadL-GBP+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E0040+J61048]
Ligation of
scFv-B[R0010+B0034+OmpA+scFv-B]
scFv-C[R0010+B0034+OmpA+scFv-C]
Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own]
Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web]
GBP[R0010+B0034+FadL-GBP+B0030+E0040+J61048]
PCR of
Anti-Her 2[Herceptin-Web]
scFv-B[J23101+B0034+OmpA+scFv-B]
scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr
Anti-Her 2[Herceptin-Own]
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar 		19

Electrophoresis of
Anti-Her 2[Herceptin-Web]
scFv-B[J23101+B0034+OmpA+scFv-B]
scFv-C[J23101+B0034+OmpA+scFv-C+pSB3C5] Cr
Anti-Her 2[Herceptin-Own]
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
scFv-C[OmpA+scFv-C]
Anti-Her 2[Herceptin-Own] (NcoI and SpeI)
Anti-Her 2[Herceptin-Web] (NcoI and SpeI)
Anti-Her 2[J23101+B0034+OmpA+Herceptin-Own] Ar (EcoRI and SpeI)
BFP[B0030+K592100+J61048] Cr (XbaI and PstI)
Transform of
GBP[R0010+B034+FadL-GBP] Ar
GBP[J23101+B0034+FadL-GBP] Ar
GBP[J23101+B0034+FadL-GBP+B0030+E0040+J61048] Cr 		20

PCR of
GBP[FadL-GBP] Kr
OmpA[OmpA] Cr
Gel extraction of
OmpA[OmpA] (EcoRI and SpeI)
Plasmid purification of
Anti-Her 2[Herceptin-Own] Cr
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar
PCR of
GBP[FadL-GBP] Kr
OmpA[R0010+B0034+OmpA+pSB1C3] Cr
GBP[R0010+B0034+GBP+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr
GBP[R0010+B0034+GBP+B0030+E0040+J61048]
scFv-B[R0010+B0034+OmpA+scFv-B]
scFv-C[R0010+B0034+OmpA+scFv-C]
Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own]
Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web]
Digestion of
scFv-C[J23101+B0034+OmpA+scFv-C+B0030+E0040+J61048+pSB6A1] Ar (EcoRI and PstI)
Anti-Her 2[Herceptin-Own] Cr
Electrophoresis of
OmpA[R0010+B0034+OmpA+pSB1C3] Cr
GBP[R0010+B0034+GBP+pSB1C3] Cr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048] Kr
GBP[R0010+B0034+GBP+B0030+E0040+J61048]
Ligation of
scFv-B[R0010+B0034+OmpA+scFv-B+pSB1K3] Kr
scFv-C[R0010+B0034+OmpA+scFv-C+pSB1K3] Kr
Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own+pSB1K3] Kr
Anti-Her 2[R0010+B0034+OmpA+Herceptin-Web+pSB1K3] Kr
GBP[R0010+B0034+GBP+B0030+E0040+J61048pSB1A3] Ar
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+E1010+J61048+pSB1K3] Kr
scFv-B[J23101+B0034+OmpA+scFv-B+B0030+K592100+J61048+pSB1K3] Kr
Anti-Her 2[R0010+B0034+OmpA+Herceptin-Own+B0030+E1010+J61048+pSB1K3] Kr 		21

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PCR
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