Team:UESTC-China/result
From 2014.igem.org
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<p style="color:#1b1b1b;">Tobacco was transformed essentially using the leaf disk co-cultivation protocol of Horsch. This protocol includes three stages, co-cultivation (Fig.4A), screening cultivation (Fig.4B) and rooting cultivation (Fig.4C). We have successfully transformed each vector into babacco. Table 1 is the statistical result of quantity of each transgenic line. And we have got PCR positive plantlet of every transgenic line.</p> | <p style="color:#1b1b1b;">Tobacco was transformed essentially using the leaf disk co-cultivation protocol of Horsch. This protocol includes three stages, co-cultivation (Fig.4A), screening cultivation (Fig.4B) and rooting cultivation (Fig.4C). We have successfully transformed each vector into babacco. Table 1 is the statistical result of quantity of each transgenic line. And we have got PCR positive plantlet of every transgenic line.</p> | ||
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- | <div align="center"><img style="width: | + | <div align="center"><img style="width:60% ;" src="https://static.igem.org/mediawiki/2014/7/75/Rr4.JPG"> |
- | <p style="position:relative; left:0px; padding:15 5px; font-size:20px; font-family: calibri, arial, helvetica, sans-serif; font-style: calibri; text-align:left; width: | + | <p style="position:relative; left:0px; padding:15 5px; font-size:20px; font-family: calibri, arial, helvetica, sans-serif; font-style: calibri; text-align:left; width:700px; color:#1b1b1b;"> |
<strong>Fig.4</strong> | <strong>Fig.4</strong> | ||
Transform piGEM003, piGEM004, piGEM005, piGEM006, piGEM007, piGEM008, piGEM009, piGEM010 and piGEM011 into tobacco. Co-cultured for 48 hours(A); Screening cultivation for one month(B); Rooting cultivation for one month(C). | Transform piGEM003, piGEM004, piGEM005, piGEM006, piGEM007, piGEM008, piGEM009, piGEM010 and piGEM011 into tobacco. Co-cultured for 48 hours(A); Screening cultivation for one month(B); Rooting cultivation for one month(C). | ||
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<h1 class="SectionTitles" style="width:1140px; ">Expression of four key enzymes in tobacco</h1><br/> | <h1 class="SectionTitles" style="width:1140px; ">Expression of four key enzymes in tobacco</h1><br/> | ||
- | <p style="color:#1b1b1b;">We extracted DNA from tobacco plantlets. And then we used specific primers to amplify the target gene to verify the kan-resistant seedlings (Fig.5). Next we extracted RNA from tobacco leaves which are PCR positive. We used RT-PCR to detect whether target gene was expressed | + | <p style="color:#1b1b1b;">We extracted DNA from tobacco plantlets. And then we used specific primers to amplify the target gene to verify the kan-resistant seedlings (Fig.5). Next we extracted RNA from tobacco leaves which are PCR positive. We used RT-PCR to detect whether target gene was expressed (Fig.6).</p> |
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<div align="center"><img style="width:50% ;" src="https://static.igem.org/mediawiki/2014/5/58/10.png"> | <div align="center"><img style="width:50% ;" src="https://static.igem.org/mediawiki/2014/5/58/10.png"> | ||
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</p> | </p> | ||
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- | <div align="center"><img style="width:10% ;" src=" | + | <div align="center"><img style="width:10% ;" src="https://static.igem.org/mediawiki/2014/4/43/RrRNA.png"> |
<p style="position:relative; left:0px; padding:15 5px; font-size:20px; font-family: calibri, arial, helvetica, sans-serif; font-style: calibri; text-align:left; width:650px; color:#1b1b1b;"><strong>Fig.6</strong> | <p style="position:relative; left:0px; padding:15 5px; font-size:20px; font-family: calibri, arial, helvetica, sans-serif; font-style: calibri; text-align:left; width:650px; color:#1b1b1b;"><strong>Fig.6</strong> | ||
- | + | <strong>Fig.6</strong>RT-PCR verification of positive transgenic tobacco seedlings.<br/> | |
+ | M: DNA marker; 1-8: 6 individual lines; 9: plasmid | ||
</p> | </p> | ||
</div> | </div> |
Revision as of 01:22, 14 October 2014