Difference between revisions of "Team:Brasil-USP/Notebook/Calendar"
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<td valign=top> 01/09 | <td valign=top> 01/09 | ||
Revision as of 05:11, 18 September 2015
Notebook
Calendar
![](https://static.igem.org/mediawiki/2015/7/7d/Team-Brasil-USP_MainLogo.png)
July
Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
01/06
- Agarose gel (RBS_hokD) - Gel purification (RBS_hokD) - Ligation 2X_RBS_hokD (RBS_hokD + RBS_hokD) - EXSP Digestion (P0040, Pl_RBS, GFP_R0440) - Agarose gel (P0040) - Gel purification (P0040) - Ligation Pt_I13507(P0040 + I13507) - Interlab gel purification and ligation 02/06 - Transformation Ptet_I13507 - Agarose gel (Pl_RBS, GFP_P0440) - Gel purification (Pl_RBS, GFP_P0440) - Ligation Pl_P0440 (Pl_RBS + GFP_P0440) - Interlab transformation (with wrong antibiotic ampicillin - is possible found in registry the same parts in another backbone BBa_J61002) |
03/06
- Transformation (Pr_RBS, 2X_RBS_hokD) - EXSP digestion (P0440, R0040) - Transformation Pl_R0440 (Pl_RBS + GFP_P0440) - Inoculum (Pt_I13507) |
04/06
- Agarose gel (P0440, R0040) - Gel purification (P0440, R0040) - Ligation tet_Ptet (P0440 + R0040) - Inoculum (2X_RBS_hokD) - Miniprep (Pt_I13507) |
05/06
- Inoculum (R0080, K91403) - EXSP digestion (tet_Term, Pt_E0840) - Miniprep (2X_RBS_hokD) |
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06/06
- NotI digestion (2X_RBS_hokD, Pr_RBS, Pa_RBS) - Transformation (tet_Ptet) - Agarose gel (2X_RBS_hokD) – positive results - Agarose gel (tet_Term, Pt_E0840) - EXSP digestion (Pt_E0480) - Inoculum (Pl_P0440) - NotI digestion (Pt_I13507) – positive results |
07/06
- Agarose gel (Pr_RBS, Pa_RBS, Pt_E0480) – positive results for Pr_RBS and Pt_E0840) - EXSP digestion (2X_RBS_hokD, B0015) - Ligation Term_E0840 (Pt_E0840 + tet_Term) - Inoculum (tet_Ptet) - Transformation (Term_E0840) - Miniprep (Pl_P0440) - Not digestion (Pl_P0440) – positive results - EXSP digestion (Pl_P0440, Pt_I13507) - Inoculum (GFP_P0440) |
08/06
- Agarose gel (2X_RBS_hokD, B0015) – B0015 didn’t digest - Gel purification (2X_RBS_hokD) - EXSP digestion (B0015) - Miniprep (tet_Ptet) - NotI digestion (tet_Ptet) - Agarose gel (tet_Ptet, B0015) – positive results - Gel digestion (B0015) - Ligation hokD_Term (2X_RBS_hokD + B0015) - Ligation Pl_I13507 (Pl_P0440 + Pt_I13507) - Miniprep (GFP_P0440) - EXSP digestion (GFP_P0440) |
09/06
- Agarose gel and gel purification (GFP_P0440) - Ligation GFP_I13507 (GFP_P0440 + Pt_I13507) - Transformation (Pl_I13507) |
10/06
- Transformation (hokD_Term) - Inoculum (Pl_I13507) - Transformation (GFP_I13507) | 11/06
- Miniprep (Pl_I13507) |
12/06 |
13/06
- Inoculum (GFP_I13507) - NotI digestion (Pl_I13507) - Agarose gel (Pl_I13507) – negative results |
14/06
- Miniprep (GFP_I13507) - NotI digestion (GFP_I13507) – negative results - EXSP digestion (Pl_P0440, Pt_I13507) - Inoculum (GFP_P0440, Pt_I13507) | 15/06
- Agarose gel (Pl_P0440, Pt_I13507) - Gel purification (Pl_P0440, Pt_I13507) - Ligation Pl_I13507 (Pl_P0440 + Pt_I13507) - Miniprep (GFP_P0440, Pt_I13507) - EXSP digestion (GFP_P0440, Pt_I13507) |
16/06
- Agarose gel and gel purification (GFP_P0440, Pt_I13507) - Ligation GFP_I13507 (GFP_P0440 + Pt_I13507) - Transformation (Pl_I13507) | 17/06
- Transformation (GFP_I13507) - Inoculum (Pt_Term, Pl_I1350) |
18/06
- Miniprep (Pt_Term, Pl_I13507) - NotI digestion (Pt_Term, Pl_I13507) - Agarose gel (Pt_Term, Pl_I13507) – positive results for Pt_Term | 19/06 |
20/06
| 21/06
- Inoculum (GFP_I13507, Pl_I13507) - New interlab transformations |
22/06
- Miniprep (GFP_I13507, Pl_I13507) - NotI digestion (GFP_I13507, Pl_I13507) - EXSP digestion (Pr_RBS, R0080) - Agarose gel (GFP_I13507, Pl_I13507, Pr_RBS) - Gel purification (Pr_RBS) - Inoculum (GFP_I13507, Pl_I13507) - Interlab inocula | 23/06
- Agarose gel (R0080) - Miniprep (GFP_I13507, Pl_I13507) - NotI digestion (GFP_I13507, Pl_I13507) - Agarose gel (GFP_I13507, Pl_I13507) – Positive results for GFP_I13507 - EXSP digestion (GFP_I13507) - Interlab miniprep and restriction analysis |
24/06
- Miniprep (B0015, Pt_P0440) - EXSP digestion (2X_RBS_hokD, B0015) - Transformation (Pa_RBS) - Agarose gel of restriction analysis: devices confirmed | 25/06
- agarose gel (2X_RBS_hokD, B0015) - B0015 did not digest |
26/06
- Agarose gel and gel purification (GFP_I13507) - Ligation Pr_I13507 (Pr_RBS + GFP_I13507) |
27/06
- EXSP digestion (Pl_RBS, Pt_Term) - Transformation (Pr_I13507, Pa_RBS) |
28/06
- Agarose gel (Pl_RBS, Pt_Term) ligation Pr_I13507(Pr_RBS + Pt_Term) |
29/06
- EXSP digestion (B0015, 2X_RBS_hokD) - agarose gel (B0015, 2X_RBS_hokD) - 2X_RBS_hokD didn’t digest - EXSP digestion overnight (2X_RBS_hokD) - transformation (Pr_Term, Pl_RBS) |
30/06
- inoculum (Pl_RBS, Pa_RBS) - agarose gel (B0015, 2X_RBS_hokD) - Digestion worked! - ligation 2X_hokD_Term (B0015 + 2X_RBS_hokD) P0440_hokD(P0440_term + 2X_RBS_hokD) |
31/06 |
August
Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
01/08 | 02/08 | |||||
03/08
- EXSP digestion (R0010, R0080, B0034, Pt_I13507) - NotI digestion (Pl_I13507) - Ligation Pa_RBS (R0080 + B0034) and Pl_RBS (R0010 + B0034) |
04/08
- NotI digestion (Pr_I13507) - Agarose gel (Pr_I13507) - Positive results!! - EXSP digestion (Pt_I13507) - Transformation (Pl_RBS and Pa_RBS) |
05/08
- agarose gel (Pl_I13507, Pt_I13507)- negative results for Pl_13507 :( - Agarose gel and gel purification (Pt_I13507) - ligation Pl_I13507 (Pl_P0440 + Pt_I13507) - inoculum (Pl_RBS, Pa_RBS) |
06/08
- Transformation (Pl_I13507, Pr_I13507) - miniprep (Pl_RBS, Pa_RBS) - NotI digestion (Pl_RBS, Pa_RBS) |
07/08
- Agarose gel (Pl_RBS, Pa_RBS) - Only Pa_RBS was positive - EXSP digestion (Pa_RBS, Pt_Term) - Ligation Pa_Term(Pa_RBS + Pt_Term) Pa_I13507 (Pa_RBS + GFP_I13507) |
08/08
- Ligation (2X_RBS_hokD + Term) - Transformation (Pa_I13507, Pa_Term) |
09/06
- Inoculum (Pa_I13507,Pl_I13507,Pa_Term) |
10/08
- miniprep (Pa_I13507, Pl_I13507, Pa_Term) - NotI digestion (Pa_I13507, Pa_Term) - Agarose gel (Pa_I13507, Pl_I13507, Pa_Term, Pl_RBS) - Positive results for Pa_I13507 and Pa_Term - Pre-inoculum of interlab study devices, BBa_K876004, and controls | 11/08
- Vector exchange digestion (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term) - Ligation with pSB1C3 (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term) - EXSP digestion (Pl_RBS, GFP_I13507, Pt_Term) - ligation (Pl_I13507, Pl_Term) - Transformation OmpA - Sample preparation of Interlab Study devices and controls (plate reader) - End-point measurement of Interlab Study devices and controls - Sample preparation of BBa_K876004 part (plate reader) - Kinetic measurement of BBa_K876004 (14 hours). |
12/08
- Transformation (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term) - Inoculum OmpA - Pre-inoculum of interlab study devices, BBa_K876004, and controls | 13/08
- Inoculum (Pr_RBS, GFP_I13507, GFP_P0440, Tet_Term) - miniprep OmpA - EXSP digestion (B0034, OmpA) - agarose gel (B0034, OmpA) - Ligation RBS_OmpA (RBS + OmpA) - Sample preparation of Interlab Study devices and controls (flow cytometry) - Measurement of Interlab Study devices and controls - Sample preparation of BBa_K876004 part (flow cytometry) - Flow cytometry measurement of BBa_K876004 (14 hours). |
14/08
- miniprep (Pr_RBS, GFP_P0440(C3), Pl_I13507) - Positive results for Pl_I13507 - and GFP_P0440 - Transformation RBS_OmpA | 15/08
- Inoculum (RBS_OmpA, GFP_I13507(C3) |
16/08
- miniprep (RBS_OmpA, GFP_I13507(C3)) - NotI digestion (RBS_OmpA, GFP_I13507(C3)) - Agarose gel (RBS_OmpA, GFP_I13507(C3)) - Ligation Pl_Term(Pl_RBS + Pl_Term) |
17/08
- Inoculum (RBS_OmpA) - Transformation (Pl_Term) |
18/08
- IDT primers and gBlocks arrived - miniprep (RBS_OmpA) - NotI digestion (RBS_OmpA) - Agarose gel (RBS_OmpA) - Postive - Vector exchange digestion (RBS_hokD, Pr_RBS) - Ligation RBS_hokD(C3), Pr_RBS(C3) - Inoculum (Pl_Term) | 19/08
- Transformation (RBS_hokD(C3), Pr_RBS(C3)) - NotI digestion (Pl_Term) - Agarose gel (Pl_Term)- positive!!! |
20/08
- gBlocks and Primers resuspension and quantification - EP digestion (Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term) - Ligation Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term with pBS1C3 - Inoculum (RBS_hokD(C3), Pr_RBS(C3)) | 21/08
- PCR for mutation of EcoRI roxA site - Agarose gel for amplification confirmation and purification of roxA PCR mutation - DpnI digestion of purificated PCR and DH5a transformation using the DNA digested - PCR for insert amplification of Lcp inserting Biobrick sites - PCR for insert amplification of Lcp inserting pETSUMO restriction sites (Lcp heterologous expression) - Agarose gel for amplification confirmation and purification of lcp PCRs - Digestion of Lcp PCR biobrick and pSB1C3 with EP - Digestion of Lcp PCR for pETSUMO and pETSUMO with restriction enzymes BamHI and HindIII - miniprep (RBS_hokD(C3), Pr_RBS(C3)) - NotI digestion (RBS_hokD(C3), Pr_RBS(C3)) - Agarose gel (RBS_hokD(C3), Pr_RBS(C3)) - All negative - Transformation (Pl_I13507, Pa_I13507, Pr_I13507, Pl_term, Pa_Term, Pr_Term) - Pre-inoculum (Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls) |
22/08
- Agarose gel purification for Lcp ligations - Transformation of Lcp for biobrick and Lcp for pETSUMO using insert and vector purified - Inoculum of 6 roxA mutant colonies - Inoculum (Pl_I13507(C3)) - Sample preparation (Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls) (plate reader) - five 96-well plates - Measurement of Pl_I13507, Pa_I13507, Pr_I13507, interlab study devices and controls (12 hours - plate reader) - Pre-inoculum (Pl_Term, Pa_Term, Pr_Term, BBa_K876004 and controls) (codes)) | 23/08
- gBlock reactions and transformations (linker-GFP, 6XHis, signalTAT) - Miniprep roxA mutants - Digestion of EcoRI and HindIII of roxA mutants and plasmid non-mutated (control) - Agarose gel of the restriction analysis - Sample preparation (Pl_Term, Pa_Term, Pr_Term, BBa_K876004 and controls) (plate reader) - five 96-well plates - Measurement of Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507 and BBa_K876004 (12 hours - plate reader) - miniprep (Pl_I13507(C3)) - NotI digestion (Pl_I13507(C3)) - Agarose gel (Pl_I13507(C3)) - posite results |
24/08
- Inoculum (linker-GFP, 6XHis, signalTAT, Lcp(pETSUMO), Lcp(pSB1C3)) - miniprep (Pa_Term, Pl_Term) - EP digestion (Pa_Term, Pr_Term, Pl_Term, 2xHokD, B0015) - PCR of mutant roxA for biobrick ligation | 25/08
- miniprep (OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)) - NotI and EXSP digestions ( (OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)) - Agarose gel ((OmpA_RBS, Pr_RBS, Pr_I13507, linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3)) - Positive results for OmpA_RBS ,linker-GFP, signalTAT, 6xHis, Lcp(pETSUMO), Lcp(pSB1C3) - Ligation 6xHis_Term (linker6xHis +B0015), GFP_Term (linkerGFP + B0015), OmpA_Lcp (OmpA + Lcp(pSB1C3)), Tat_Lcp (signalTAT + Lcp(pSB1C3) - Ligation (Pa_Term(C3), Pl_Term(C3), Pr_Term(C3) - Agarose gel (mutant RoxA) |
26/08
- Transformation 6xHis_Term, GFP_Term, OmpA_Lcp, Tat_Lcp | 27/08
- Inoculum (6xHis_Term, GFP_Term, mpA_Lcp, Tat_Lcp) - Transformation (2xhokD_Term) - PCR of mutant roxA for biobrick ligation - Agarose gel of mutant roxA PCR product |
28/08
- roxA PCR product purification - roxA PCR product digestion with EP - miniprep (6xHis_Term, GFP_Term, OmpA_Lcp, Tat_Lcp) - NotI and EXSP digestions (6xHis_Term, GFP_Term, OmpA_Lcp, Tat_Lcp) - Only 6xHis_Term was negative - ligation Lcp_Term I (Tat_Lcp + GFP_Term), Lcp_Term II (OmpA_Lcp + GFP_Term) - Pre-inoculum (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507) |
29/08
- Agarose gel (roxA PCR product) - Ligation roxA(mutant) +pSB1C3 - Sample preparation (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507) (plate reader) - seven 96-well plates - Measurement of test promoters (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507) |
30/08
- Transformation (roxA) |
31/08
- Inoculum (roxA) - Transformation (Lcp_Term I, Lcp_Term II) |
September
Monday | Tuesday | Wednesday | Thursday | Friday | Saturday | Sunday |
01/09
- miniprep (roxA) - Inoculum (Lcp_Term I) - Ligation (Lcp_Term II) |
02/09
- miniprep (Lcp_Term I) - digestion (Lcp_Term I, roxA)) - Transformation (Lcp_Term II) |
03/09
- Transformation (Pr_RBS, 2X_RBS_hokDLigation Pl_hokD (Pl + 2xhokD)) -Pre-inoculum (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507) |
04/09
- Transformation (Pl_2xhokD) -Sample preparation of test promoters circuits (plate reader) - Measurement of test promoters circuits (Pl_Term, Pa_Term, Pr_Term, Pl_I13507, Pa_I13507, Pr_I13507) |
05/09
- Inoculum (Pl_2xhokD) |
06/09
- Transformation in Bl21(DE3) (Pl_I13507, Pa_I13507, Pr_I13507, Pl_Term, Pa_Term, Pr_Term) |
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07/09
- Inoculum (Pl_I13507, Pa_I13507, Pr_I13507, Pl_Term, Pa_Term, Pr_Term, in BL21(DE3)) |
08/09
- Miniprep (Pl_I13507, Pa_I13507, Pr_I13507, Pl_Term, Pa_Term, Pr_Term, in BL21(DE3)) - Digestion with SacI (OmpA_Lcp, OmpA_roxA, TAT_Lcp, TAT_roxA) - Digestion with SacI and XbaI (OmpA, signalTAT, Lcp, RoxA) - EXSP digestion (R0010, R0080, K914003) -NotI digestion (Pl_I13507, Pa_I13507, Pr_I13507, Pl_Term, Pa_Term, Pr_Term, in BL21(DE3)) - Agarose gel (OmpA_Lcp, OmpA_roxA, TAT_Lcp, TAT_roxA, OmpA, signalTAT, Lcp, RoxA, Pl_I13507, Pa_I13507, Pr_I13507, Pl_Term, Pa_Term, Pr_Term) - Positive results for Pl_I13507 and Pr_I13507) - Ligation OmpA_Lcp, OmpA_roxA, TAT_Lcp, TAT_roxA, Pl_I13507II (R0010 + GFP_I13507), Pl_TermII (R0010 + tet_Term), Pa_I13507II (R0080 + GFP_I13507), Pa_TermII (R0080 + tet_Term), Pr_I13507II (K914003 + GFP_I13507), Pr_TermII (K914003 + tet_Term), -Transformation (OmpA_Lcp, OmpA_roxA, TAT_Lcp, TAT_roxA, Pl_I13507II, Pa_I13507II, Pr_I13507II, Pl_TermII, Pa_TermII, Pr_TermII) |
09/09
-Western blot to see if LCP was been expressed in pETSUMO - inoculum (Pl_I13507II, Pa_I13507II, Pl_TermII, Pa_TermII, Pr_TermII, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt) - Prepare plates for characterization (Pl_I13507 and Pr_I13507 in BL21(DE3)) 10th |
10/09
- Miniprep (Pl_I13507II, Pa_I13507II, Pl_TermII, Pa_TermII, Pr_TermII, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt) -NotI digestion (Pl_I13507II, Pe_I13507II, Pl_TermII, Pa_TermII, Pr_TermII) - SacI digestion again (OmpA_Lcp, OmpA_roxA, TAT_Lcp, TAT_roxA, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt) -Agarose gel (Pl_I13507II, Pe_I13507II, Pl_TermII, Pa_TermII, Pr_TermII, OmpA_Lcp, OmpA_roxA, TAT_Lcp, TAT_roxA, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt) -Ligation (OmpA_Lcp, OmpA_roxA, TAT_Lcp, TAT_roxA, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt, GFP_TermA (J23101 + linkerGFP_Term), GFP_TermB (J23117 + linkerGFP_Term), GFP_TermC (J23106 + linkerGFP_Term), -Transformation (OmpA_Lcp, OmpA_roxA, TAT_Lcp, TAT_roxA, TAT_Lcp_RBS, OmpA_Lcp_RBS, OmpA_roxA_Pt, GFP_TermA, GFP_TermB, GFP_TermC | 11/09
- Transformation (TAT_ lcp_RBS, TAT_ lcp, TAT_ roxA, OmpA_roxA_Pt) -Inoculum (GFP_TermA, GFP_TermB, GFP_TermC) |
12/09
- Miniprep (GFP_TermA, GFP_TermB, GFP_TermC) - Not digestion (GFP_TermA, GFP_TermB, GFP_TermC) - Xbal and NotI digestion (RoxA, signalTAT and Lcp) -Agarose gel (GFP_TermA, GFP_TermB, GFP_TermC, RoxA, signalTAT and Lcp) -Ligation TAT_Lcp (signalTAT + Lcp), TAT_RoxA (signalTAT + roxA) | 13/09
- Transformation (TAT_Lcp, TAT_roxA) |
14/09
- EP digestion to change the vector (Pa_Term, signalTAT, OmpA, Lcp, RoxA) - SacI and EcoRI digestion (signalTat, OmpA) - SacI and PstI digestion (Lcp, RoxA) - Agarose gel ( Pa_Term, signalTAT, OmpA, Lcp, RoxA) -Ligation (pSB1A2 + RoxA, pSB1A2 + Lcp, pSB1A2 + OmpA and pSB1A2 + SignalTAT) -3A assembly (TAT_RoxA(pSB1A2 +signalTA+RoxA), TAT_Lcp(pSB1A2 +signalTA+Lcp), OmpA_RoxA(pSB1A2 +OmpA+RoxA),OmpA_Lcp( pSB1A2 +OmpA+Lcp)) -Transformation (RoxA, Lcp, OmpA, SignalTAT, TAT_RoxA, TAT_Lcp, OmpA_RoxA, OmpA_Lcp) - Inoculum (linkerGFP_J23117, linkerGFP_J23106) | 15/09
- Ligation again (pSB1A2 + RoxA, pSB1A2 + Lcp, pSB1A2 + OmpA and pSB1A2 + SignalTAT) - 3A assembly again (TAT_RoxA(pSB1A2 +signalTA+RoxA), TAT_Lcp(pSB1A2 +signalTA+Lcp), OmpA_RoxA(pSB1A2 +OmpA+RoxA),OmpA_Lcp( pSB1A2 +OmpA+Lcp)) - Sample preparation (linkerGFP_J23117, linkerGFP_J23106) - Measurements (linkerGFP_J23117, linkerGFP_J23106) |
16/09
- Transformation (RoxA, Lcp, OmpA, SignalTAT, TAT_RoxA, TAT_Lcp, OmpA_RoxA, OmpA_Lcp) - Prepare the plates to submit the new parts | 17/09
- Inoculum (RoxA, Lcp, OmpA, SignalTAT, TAT_RoxA, TAT_Lcp, OmpA_RoxA, OmpA_Lcp) |
18/09
- Miniprep (RoxA, Lcp, OmpA, SignalTAT, TAT_RoxA, TAT_Lcp, OmpA_RoxA, OmpA_Lcp) - NotI digestion (RoxA, Lcp, OmpA, SignalTAT, TAT_RoxA, TAT_Lcp, OmpA_RoxA, OmpA_Lcp) - Agarose gel (RoxA, Lcp, OmpA, SignalTAT, TAT_RoxA, TAT_Lcp, OmpA_RoxA, OmpA_Lcp) |