Difference between revisions of "Part:BBa K3376000"
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Lactate dehydrogenase catalyzes the fermentation of pyruvate to lactic acid. It plays a pivotal role in the glucose metabolic pathway connecting glycolysis, and is recognized as an essential gene and expressed at high level in S. mutans. Its promoter was cloned out from the gDNA of S. mutans and tested in transformed E. coli. | Lactate dehydrogenase catalyzes the fermentation of pyruvate to lactic acid. It plays a pivotal role in the glucose metabolic pathway connecting glycolysis, and is recognized as an essential gene and expressed at high level in S. mutans. Its promoter was cloned out from the gDNA of S. mutans and tested in transformed E. coli. | ||
| + | === Expression === | ||
| + | The fluorescence expression levels were measured for lactate dehydrogenase promoter (ldhp) [ldhp-GFP-Tr/pSB1C3 (BBa_K3376002)] and thiol peroxidase promoter (tpxp) [tpxp-GFP-Tr/pSB1C3 (BBa_K3376004)] activity of S. mutans. GFP was detected at Ex/Em = 483/513. As shown in the figure, the expression level of ldhp-driven GFP was very high compared to a regulated promoter (tpxp) and a vector-only control. | ||
| + | [[File:T--Mingdao--ww1.png|450px|center]] | ||
| + | |||
| + | === Transformation === | ||
| + | The ldhp-GFP-Tr/pSB1C3 [BBa_K3376002] was transformed into E. coli Nissle strain. The high expression of green fluorescent protein was observed under a blue led light. | ||
| + | |||
| + | [[File:T--Mingdao--ww2.png|600px|center]] | ||
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Revision as of 15:24, 6 October 2020
S. mutans lactate dehydrogenase promoter (ldhp)
Lactate dehydrogenase catalyzes the fermentation of pyruvate to lactic acid. It plays a pivotal role in the glucose metabolic pathway connecting glycolysis, and is recognized as an essential gene and expressed at high level in S. mutans. Its promoter was cloned out from the gDNA of S. mutans and tested in transformed E. coli.
Expression
The fluorescence expression levels were measured for lactate dehydrogenase promoter (ldhp) [ldhp-GFP-Tr/pSB1C3 (BBa_K3376002)] and thiol peroxidase promoter (tpxp) [tpxp-GFP-Tr/pSB1C3 (BBa_K3376004)] activity of S. mutans. GFP was detected at Ex/Em = 483/513. As shown in the figure, the expression level of ldhp-driven GFP was very high compared to a regulated promoter (tpxp) and a vector-only control.
Transformation
The ldhp-GFP-Tr/pSB1C3 [BBa_K3376002] was transformed into E. coli Nissle strain. The high expression of green fluorescent protein was observed under a blue led light.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]