Difference between revisions of "Part:BBa K1675019:Design"
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===References=== | ===References=== | ||
| + | ［1］ Le Y, Chen H, Zagursky R, et al. Thermostable DNA ligase-mediated PCR production of circular plasmid (PPCP) and its application in directed evolution via in situ error-prone PCR[J]. DNA research, 2013: dst016. | ||
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| + | ［2］ Pai J C, Entzminger K C, Maynard J A. Restriction enzyme-free construction of random gene mutagenesis libraries in Escherichia coli[J]. Analytical biochemistry, 2012, 421(2): 640-648. | ||
Latest revision as of 00:08, 28 September 2015
P-atp2 mutant 202-B0034-lacz alpha
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
There is an EcoRI site exists in the P-atp2 sequence, so we mutate it to change it.
Source
The natural P-atp2 is an alkali-induced promoter in Corynebacterium glutamicum located in F0F1 ATPase operon. The natural P-atp2 promoter responds to the pH changes from pH 7.0 to pH 9.0, especially at alkaline pH. When the pH value increasing, the expression increased modestly. It is activated by the alternative sigma factor of the RNA polymerase, whose synthesis would be activated when the bacteria are growing at alkaline pH.
The activity of this mutant at different pH is shown in figure 1.
References
［1］ Le Y, Chen H, Zagursky R, et al. Thermostable DNA ligase-mediated PCR production of circular plasmid (PPCP) and its application in directed evolution via in situ error-prone PCR[J]. DNA research, 2013: dst016.
［2］ Pai J C, Entzminger K C, Maynard J A. Restriction enzyme-free construction of random gene mutagenesis libraries in Escherichia coli[J]. Analytical biochemistry, 2012, 421(2): 640-648.