Team:Caltech/Notebook/Degradation
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<font size="+2"><a name="7_16_20_12">July 16-20, 2012</a></font> | <font size="+2"><a name="7_16_20_12">July 16-20, 2012</a></font> | ||
+ | <br> The pMQ30 is a deletion plasmid. We are using this plasmid to knock out expression of NADH dehydrogenase. According to the paper ''Respiratory chain analysis of high ethanol producing Zymomonas mobilis mutants,'' a mutant strain of Z. mobilis lacking NADH dehydrogenase had increased ethanol production. However, pMQ30 has gentamicin resistance, and Zymomonas is already resistant to gentamicin, so we are replacing it with tetracycline resistance from pSB1T3. | ||
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+ | <br> The pLAFR5 plasmid is being used for conjugation from E. coli into Z. mobilis. | ||
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+ | <br> DKN79 (DH5alpha + pLAFR5) has been plated to grow more copies of pLAFR5. | ||
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+ | <br> DKN1005 (BW20427 [WM3064] + pLAFR5) has been plated to be used for testing conjugation into Z. mobilis ZM4. | ||
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+ | <br> The pMQ95 and pMQ97 plasmids Zymomonas | ||
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<br> Electroporated pSB1T3 into competent E coli cells, incubated in LB for an hour, and streaked transformants onto tetra plates | <br> Electroporated pSB1T3 into competent E coli cells, incubated in LB for an hour, and streaked transformants onto tetra plates | ||
- | <br> Streaked DH5alpha | + | <br> Streaked DH5alpha + pMQ30 onto a gentamicin 20 ug/mL plate |
<br> Streaked Z. mobilis ZM4 from ATCC onto a RM plate and liquid culture. | <br> Streaked Z. mobilis ZM4 from ATCC onto a RM plate and liquid culture. | ||
<br> Streaked out DKN79 (DH5alpha + pLAFR5) onto a tetracycline plate | <br> Streaked out DKN79 (DH5alpha + pLAFR5) onto a tetracycline plate | ||
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<br> Made LB media | <br> Made LB media | ||
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+ | <br> No growth with pSB1T3 transformants, so retransformed and plated E coli + pSB1T3 | ||
+ | <br> Kept a liquid culture of E coli + pSB1T3 | ||
+ | <br> Ordered primers as part of switching out the pSB1T3 plasmid's resistance to tetracycline | ||
+ | <br> Z. mobilis plate has no growth | ||
+ | <br> Sent DNAWorks requests for als genes (alsBACE) | ||
+ | <br> Re-streaked out DH5alpha + pMQ30 because first plate's growth was poor | ||
+ | <br> Streaked out pMQ95 | ||
+ | <br> Streaked out pMQ97 | ||
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<a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | <a a style="float:right;" href="https://2012.igem.org/Team:Caltech/Notebook/Degradation#Calendar">Back to Degradation Notebook Calendar</a> | ||
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Revision as of 18:29, 18 July 2012
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Degradation Notebook
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