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| − | <p class=MsoNormal><b><span lang=EN-US>Background<o:p></o:p></span></b></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>Yogurt, as one of the oldest and most
| + | |
| − | ordinary fermented foods, is known all of the world. However, the yogurt
| + | |
| − | section practically taken over the dairy aisle of most grocery stores. Differ
| + | |
| − | from other traditional dairy products(cottage cheese, sour cream and etc.),
| + | |
| − | yogurt wins itself the most popularity due to the many health benefits that it
| + | |
| − | can gives.</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>And, what kinds of health benefits of
| + | |
| − | yogurt?</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>Primarily, everyone needs to have a healthy
| + | |
| − | amount of ''good'' bacteria in the digestive tract, and many yogurts are made
| + | |
| − | using active, good bacteria. And then, let us notice that yogurt comes from
| + | |
| − | milk and is as well nutritionally rich in protein, calcium, riboflavin, vitamin
| + | |
| − | B6 and vitamin B16, but it has nutritional benefits beyond those of milk, namely
| + | |
| − | due to its probiotics. In fact, the health benefits of yogurt are so impressive
| + | |
| − | that many health-conscious people make it a daily habit. Here are five possible
| + | |
| − | health benefits of having a yogurt a day: </span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(1) Yogurt may help prevent Osteoporosis,
| + | |
| − | </span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(2) Yogurt may reduce the risk of high
| + | |
| − | blood pressure.</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(3) Yogurt with active cultures helps the
| + | |
| − | gut</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(4) Yogurt with active cultures may
| + | |
| − | discourage vaginal infections </span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(5) Yogurt may help you feel fuller </span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>In spite of this health benefits, spoiled
| + | |
| − | yogurt is more harm than good if you carelessly eat it. Firstly, let us talk
| + | |
| − | about the shelf-life of yogurt. Yogurt provides two significant barriers to
| + | |
| − | pathogen growth (heat and acidity) to ensure a safe product. Discard batches
| + | |
| − | that fail to set properly, especially those due to culture errors. Yogurt
| + | |
| − | generally has a 10-21 day self-life when made and stored properly in the
| + | |
| − | refrigerator below 40¡ãF. Then let we look into yogurt manufacturing process:</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(1) pasteurization</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(2) combine ingredients</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(3) inoculate</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(3) incubate</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span>(4) refrigerate</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>The first step pasteurization is used for
| + | |
| − | sterilization, but the pathogenic bacteria like <i>E.coli</i>, <i>salmonella</i>
| + | |
| − | and<i> bacillus</i> still possibly survive from the first step that can spoil
| + | |
| − | the yogurt during prolonged storage. </span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>However, from the view of the premise of
| + | |
| − | self-life is numerous and strict, is self-life really reliable? On the one
| + | |
| − | hand, there are too many reasons like long-time transport can not make sure
| + | |
| − | yogurt is stored in a 40¡ãF environment. On the other hand, when you do yogurt yourself,
| + | |
| − | you should also need to make sure the use of clean equipment and containers. To
| + | |
| − | make matter worse, if yogurt spoiled within expiration date you can not tell
| + | |
| − | them apart, for their appearances were almost the same.(illustrated by fig.1)</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><!--[if gte vml 1]><v:shapetype id="_x0000_t75"
| + | |
| − | coordsize="21600,21600" o:spt="75" o:preferrelative="t" path="m@4@5l@4@11@9@11@9@5xe"
| + | |
| − | filled="f" stroked="f">
| + | |
| − | <v:stroke joinstyle="miter"/>
| + | |
| − | <v:formulas>
| + | |
| − | <v:f eqn="if lineDrawn pixelLineWidth 0"/>
| + | |
| − | <v:f eqn="sum @0 1 0"/>
| + | |
| − | <v:f eqn="sum 0 0 @1"/>
| + | |
| − | <v:f eqn="prod @2 1 2"/>
| + | |
| − | <v:f eqn="prod @3 21600 pixelWidth"/>
| + | |
| − | <v:f eqn="prod @3 21600 pixelHeight"/>
| + | |
| − | <v:f eqn="sum @0 0 1"/>
| + | |
| − | <v:f eqn="prod @6 1 2"/>
| + | |
| − | <v:f eqn="prod @7 21600 pixelWidth"/>
| + | |
| − | <v:f eqn="sum @8 21600 0"/>
| + | |
| − | <v:f eqn="prod @7 21600 pixelHeight"/>
| + | |
| − | <v:f eqn="sum @10 21600 0"/>
| + | |
| − | </v:formulas>
| + | |
| − | <v:path o:extrusionok="f" gradientshapeok="t" o:connecttype="rect"/>
| + | |
| − | <o:lock v:ext="edit" aspectratio="t"/>
| + | |
| − | </v:shapetype><v:shape id="ͼƬ_x0020_1" o:spid="_x0000_i1025" type="#_x0000_t75"
| + | |
| − | style='width:334.8pt;height:222.6pt;mso-wrap-style:square;
| + | |
| − | mso-position-horizontal-relative:page;mso-position-vertical-relative:page'>
| + | |
| − | <v:imagedata src="Wiki-----Project_files/image001.png" o:title="" croptop="8304f"
| + | |
| − | cropright="2121f"/>
| + | |
| − | </v:shape><![endif]--><![if !vml]><img width=558 height=371
| + | |
| − | src="https://static.igem.org/mediawiki/2015/f/f1/NEFU_FI_TEST-1.png" v:shapes="ͼƬ_x0020_1"><![endif]><span
| + | |
| − | style='mso-tab-count:1'> </span></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>fig.1(those nine photos of yogurt with a
| + | |
| − | same production date were taken maintain a two days interval. Those yogurt were
| + | |
| − | kept in room temperature, and their packing were open only when took photos )</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><span
| + | |
| − | style='mso-spacerun:yes'> </span></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>This is for sure, the pathogenic bacteria
| + | |
| − | can be detected. We then did an experiment based on our national food safety
| + | |
| − | standards. (<u>protocols click here</u>) </span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>Needless to say, no one would willing to
| + | |
| − | eat yogurt with such amount of coliform if he or she had see our results.
| + | |
| − | However, considering the test method containing cumbersome steps, costing much
| + | |
| − | time and requiring certain experimental apparatus, it on no condition can be
| + | |
| − | use in daily life. That is to say, if we don¡¯t do something we could still eat
| + | |
| − | yogurt that maybe contain much bacteria we don¡¯t want.</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US>Now, a handy detector of pathogenic
| + | |
| − | bacteria is really needed.</span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><b><span lang=EN-US style='mso-bidi-font-size:10.5pt'>Design:<o:p></o:p></span></b></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-left:0cm;text-indent:0cm;mso-list:l0 level1 lfo1'><![if !supportLists]><span
| + | |
| − | lang=EN-US style='mso-bidi-font-size:10.5pt;mso-fareast-font-family:"Times New Roman"'><span
| + | |
| − | style='mso-list:Ignore'>1.</span></span><![endif]><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>Introduction of AI-2<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'>2.Why we
| + | |
| − | choose AI-2<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'>3.Chassis(why
| + | |
| − | we choose them)<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'>4.The
| + | |
| − | system we will construct<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'>5.NICE
| + | |
| − | system<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'>6.Plasmid
| + | |
| − | construction<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'>7.Quantification<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><span
| + | |
| − | style='mso-spacerun:yes'> </span>Quorum sensing (QS) is a chemical communication
| + | |
| − | system among bacterium,and QS requires the production and detection of specific
| + | |
| − | signaling molecules,which are commonly referred to as
| + | |
| − | autoinducers.Autoinducer-2 (AI-2) molecules are one class of signaling
| + | |
| − | molecules involved in QS.AI-2 is a byproduct of the Activated Methyl
| + | |
| − | Cycle,which recycles S-adenosyl-l-methionine (SAM).As a main methyl donor,SAM
| + | |
| − | is converted to S-adenosyl-l-homocysteine (SAH).Then SAH is subsequently
| + | |
| − | hydrolyzed to adenine and 4,5-dihydroxy-2,3-pentanedione (DPD) by the nucleosidase
| + | |
| − | Pfs and the Luxs enzyme.Upon formation,DPD spontaneously cyclizes to form at
| + | |
| − | least two different interspecies communication molecules described as AI-2.<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><span
| + | |
| − | style='mso-spacerun:yes'> </span>Why we chose AI-2 as signaling
| + | |
| − | molecule,that¡¯s because the production of AI-2 is the only quorum-sensing
| + | |
| − | mechanism shared by both Gram-positive and Gram-negative bacteria.There are
| + | |
| − | many <a name="OLE_LINK7"></a><a name="OLE_LINK6"><span style='mso-bookmark:
| + | |
| − | OLE_LINK7'>pathogens</span></a> in spoiled yogurt, Ecoli, Salmonella and
| + | |
| − | Bacillus are the main part.Then AI-2 can be used for a universal language among
| + | |
| − | pathogens.Besides,AI-2 response pathway of our chassis, lactobacillus and
| + | |
| − | Lactococcus, is different from that kinds of pathogens.<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'><span style='mso-spacerun:yes'>
| + | |
| − | </span>We choose lactobacillus and Lactococcus as our chassis.As beneficial
| + | |
| − | bacteria, they are food grade and are widely used in food production for
| + | |
| − | thousands of years.Lactobacillus can improve nutrient absorption and human
| + | |
| − | gastrointestinal function.They can also inhibit the reproduction of pathogenic
| + | |
| − | bacteria in guts. Lactococcus lactis is a Gram-positive lactic acid bacterium
| + | |
| − | that is widely used in food fermentations, such as in cheese and butter
| + | |
| − | production. <o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'><span style='mso-spacerun:yes'>
| + | |
| − | </span>LuxS converts SRH to the precursor of AI-2,DPD,in the cytoplasm.DPD then
| + | |
| − | undergoes spontaneous cyclization and expert to the culture
| + | |
| − | supernatant.Depending on the bacteria,response to AI-2 can follow one of the
| + | |
| − | two currently identified routes.In pathogens exemplified by Salmonella, AI-2 is
| + | |
| − | bound to a homologue of the periplasmic binding protein LsrB.LsrB is part of an
| + | |
| − | ABC transporter(encoded by the lsrACDBRK operon).This operon is regulated by
| + | |
| − | the repressor LsrR,the gene of which is located upstream. Immediately
| + | |
| − | downstream of lsrR is a gene (lsrK). LsrK phosphorylates AI-2, Phosphorylated
| + | |
| − | AI-2 then causes LsrR to relieve its repression of the lsr operon, allowing
| + | |
| − | further AI-2 import. <o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'><span style='mso-spacerun:yes'> </span>The
| + | |
| − | nisin-controlled gene expression system NICE of Lactococcus lactis is one of
| + | |
| − | the most widely used expression systems in Gram-positive bacteria.Biosynthesis
| + | |
| − | of nisin is encoded by a cluster of 11 genes, NisR and NisK belong to the
| + | |
| − | family of bacterial two-component signal transduction systems.NisK is a
| + | |
| − | histidine</span><span style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;
| + | |
| − | mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:"Times New Roman"'>–</span><span
| + | |
| − | lang=EN-US style='mso-bidi-font-size:10.5pt'>protein kinase that resides in the
| + | |
| − | cytoplasmic membrane and is proposed to act as a receptor for the mature nisin
| + | |
| − | molecule. Upon binding of nisin to NisK, it autophosphorylates and transfers the
| + | |
| − | phosphate group to NisR, which is a response regulator that becomes activated
| + | |
| − | upon phosphorylation by NisK. Activated NisR* induces transcription from two of
| + | |
| − | the three promoters in the nisin gene cluster: PnisA and PnisF. In our parts,we
| + | |
| − | use inducible promoter PnisA to express gene A</span><span style='mso-bidi-font-size:
| + | |
| − | 10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:
| + | |
| − | "Times New Roman"'>¡¢</span><span lang=EN-US style='mso-bidi-font-size:10.5pt'>B</span><span
| + | |
| − | style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";
| + | |
| − | mso-hansi-font-family:"Times New Roman"'>¡¢</span><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>C</span><span style='mso-bidi-font-size:10.5pt;
| + | |
| − | font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:
| + | |
| − | "Times New Roman"'>¡¢</span><span lang=EN-US style='mso-bidi-font-size:10.5pt'>D</span><span
| + | |
| − | style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";
| + | |
| − | mso-hansi-font-family:"Times New Roman"'>¡¢</span><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>R</span><span style='mso-bidi-font-size:10.5pt;
| + | |
| − | font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:
| + | |
| − | "Times New Roman"'>¡¢</span><span lang=EN-US style='mso-bidi-font-size:10.5pt'>K,the
| + | |
| − | inducer nisin</span><span style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;
| + | |
| − | mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:"Times New Roman"'>¡¢</span><span
| + | |
| − | lang=EN-US style='mso-bidi-font-size:10.5pt'>gene nisR</span><span
| + | |
| − | style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";
| + | |
| − | mso-hansi-font-family:"Times New Roman"'>¡¢</span><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>nisK are necessary for PnisA.<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><o:p> </o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'>Plasmid
| + | |
| − | construction<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'>¢ÙGene<i>
| + | |
| − | A</i></span><i><span style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;
| + | |
| − | mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:"Times New Roman"'>¡¢</span></i><i><span
| + | |
| − | lang=EN-US style='mso-bidi-font-size:10.5pt'>B</span></i><i><span
| + | |
| − | style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";
| + | |
| − | mso-hansi-font-family:"Times New Roman"'>¡¢</span></i><i><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>C</span></i><i><span style='mso-bidi-font-size:
| + | |
| − | 10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:
| + | |
| − | "Times New Roman"'>¡¢</span></i><i><span lang=EN-US style='mso-bidi-font-size:
| + | |
| − | 10.5pt'>D</span></i><i><span style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;
| + | |
| − | mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:"Times New Roman"'>¡¢</span></i><i><span
| + | |
| − | lang=EN-US style='mso-bidi-font-size:10.5pt'>R</span></i><i><span
| + | |
| − | style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";
| + | |
| − | mso-hansi-font-family:"Times New Roman"'>¡¢</span></i><i><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>K </span></i><i><span style='mso-bidi-font-size:
| + | |
| − | 10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:
| + | |
| − | "Times New Roman"'>¡¢</span></i><i><span lang=EN-US style='mso-bidi-font-size:
| + | |
| − | 10.5pt'>plsr </span></i><span lang=EN-US style='mso-bidi-font-size:10.5pt'>was
| + | |
| − | cloned from Salmonella typhimurium<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'>¢ÚPlasmid
| + | |
| − | pNZ8148 served for amplifying gene<i> B</i></span><i><span style='mso-bidi-font-size:
| + | |
| − | 10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:
| + | |
| − | "Times New Roman"'>¡¢</span></i><i><span lang=EN-US style='mso-bidi-font-size:
| + | |
| − | 10.5pt'>R</span></i><i><span style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;
| + | |
| − | mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:"Times New Roman"'>¡¢</span></i><i><span
| + | |
| − | lang=EN-US style='mso-bidi-font-size:10.5pt'>K </span></i><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>respectively<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><span
| + | |
| − | style='mso-spacerun:yes'> </span>Plasmid pBBR1MCS-5 served for amplifying
| + | |
| − | gene<i> A</i></span><i><span style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;
| + | |
| − | mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:"Times New Roman"'>¡¢</span></i><i><span
| + | |
| − | lang=EN-US style='mso-bidi-font-size:10.5pt'>C</span></i><i><span
| + | |
| − | style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";
| + | |
| − | mso-hansi-font-family:"Times New Roman"'>¡¢</span></i><i><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>D </span></i><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>respectively<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'><span style='mso-spacerun:yes'>
| + | |
| − | </span>We replace promoter of pET-14b with <i>plsr</i>,The final construct
| + | |
| − | called pHY300PLK contains a<span
| + | |
| − | style='mso-spacerun:yes'> </span>blue pigment gene.<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>¢ÛPlasmid pNZ8148(¡Á3)</span><span
| + | |
| − | style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";
| + | |
| − | mso-hansi-font-family:"Times New Roman"'>¡¢</span><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>pBBR1MCS-5(¡Á3) and pHY300PLK were linearized
| + | |
| − | and introduced by electroporation into our chassis.<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>¢ÜAs for eletroporation,we have two different
| + | |
| − | routes.One of which is that seven linearized palsmid and pNZ9530(Low copy
| + | |
| − | plasmid with pAM¦Â1 origin of replication, which carries the nisR and nisK
| + | |
| − | genes.) were introduced into lactobacillus.Or seven linearized palsmid containing
| + | |
| − | gene A</span><span style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:
| + | |
| − | "Times New Roman";mso-hansi-font-family:"Times New Roman"'>¡¢</span><span
| + | |
| − | lang=EN-US style='mso-bidi-font-size:10.5pt'>B</span><span style='mso-bidi-font-size:
| + | |
| − | 10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:
| + | |
| − | "Times New Roman"'>¡¢</span><span lang=EN-US style='mso-bidi-font-size:10.5pt'>C</span><span
| + | |
| − | style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";
| + | |
| − | mso-hansi-font-family:"Times New Roman"'>¡¢</span><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>D</span><span style='mso-bidi-font-size:10.5pt;
| + | |
| − | font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:
| + | |
| − | "Times New Roman"'>¡¢</span><span lang=EN-US style='mso-bidi-font-size:10.5pt'>R</span><span
| + | |
| − | style='mso-bidi-font-size:10.5pt;font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";
| + | |
| − | mso-hansi-font-family:"Times New Roman"'>¡¢</span><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'>K</span><span style='mso-bidi-font-size:10.5pt;
| + | |
| − | font-family:ËÎÌå;mso-ascii-font-family:"Times New Roman";mso-hansi-font-family:
| + | |
| − | "Times New Roman"'>¡¢</span><span lang=EN-US style='mso-bidi-font-size:10.5pt'>Blue
| + | |
| − | pigment were introduced into Lactococcus NZ9000(containing nisR and nisK)by eletroporation.<o:p></o:p></span></p>
| + | |
| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US style='mso-bidi-font-size:10.5pt'><span
| + | |
| − | style='mso-spacerun:yes'> </span><o:p></o:p></span></p>
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| − | | + | |
| − | <p class=MsoNormal style='margin-bottom:12.0pt'><span lang=EN-US
| + | |
| − | style='mso-bidi-font-size:10.5pt'><span style='mso-spacerun:yes'>
| + | |
| − | </span>The molecular structure of AI-2 is somewhat similar to ascorbic acid. It
| + | |
| − | was expected that AI-2 would also act as a reducing agent and reduce Fe(III)
| + | |
| − | ions in the presence of 1,10-phenanthroline to form the colored [(o-phen)3
| + | |
| − | Fe(II)]SO4 ferroin complex that could be quantified colorimetrically.<o:p></o:p></span></p>
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| − | | + | |
| − | <p class=MsoNormal><span lang=EN-US><o:p> </o:p></span></p>
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| − | | + | |
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