Difference between revisions of "Part:BBa K4183009"
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lacZ and kfiC share the BBa_J23119 promoter to construct acid-tolerant lactose-degrading engineered bacteria. | lacZ and kfiC share the BBa_J23119 promoter to construct acid-tolerant lactose-degrading engineered bacteria. | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
| + | To improve the transcription rate of the expression frame and to reduce the use of IPTG in humans, we replaced BBa_R0010 with BBa_J23119 and deleted the self-contained lacI element on pET28a, constructed a new complex element, BBa_K4183009, and transferred it into EcN, thus developing the second generation engineered bacterium lacMAN2.0. | ||
| + | We then performed the same experiment on it. The results showed that the lacZ secretion efficiency of lacMAN2.0 was somewhat improved to 14.49% relative to lacMAN1.0, but still lower than the initial 21.93%. In terms of acid tolerance, lacMAN2.0 could grow normally in the first 30 min in an acidic environment with pH ≤ 4, consistent with that in a normal environment. | ||
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| + | [[File:HS_China_engineering-10.png.jpg|900px|thumb|none|Figure1. Measurement of enzyme activity and acid resistance of LacMAN 2.0 | ||
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| + | (A) Determination of enzymatic activity of lacZ in lacMAN2.0 in the bacteriophage as well as in the supernatant using the kit. | ||
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| + | (B) Determination of the growth of lacMAN2.0 in acidic environments with different pH.]] | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Revision as of 15:05, 11 October 2022
Composite element of lacZ and kfiC
lacZ and kfiC share the BBa_J23119 promoter to construct acid-tolerant lactose-degrading engineered bacteria.
Usage and Biology
To improve the transcription rate of the expression frame and to reduce the use of IPTG in humans, we replaced BBa_R0010 with BBa_J23119 and deleted the self-contained lacI element on pET28a, constructed a new complex element, BBa_K4183009, and transferred it into EcN, thus developing the second generation engineered bacterium lacMAN2.0.
We then performed the same experiment on it. The results showed that the lacZ secretion efficiency of lacMAN2.0 was somewhat improved to 14.49% relative to lacMAN1.0, but still lower than the initial 21.93%. In terms of acid tolerance, lacMAN2.0 could grow normally in the first 30 min in an acidic environment with pH ≤ 4, consistent with that in a normal environment.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]