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A kind of isoprenoid transferase comq mutant, gene, vector, engineering bacteria and preparation method and application

An isoprenoid and transferase technology, which is applied in the field of molecular biology to achieve the effect of improving biosynthesis capacity and reducing protein expression level

Active Publication Date: 2022-03-08
ANHUI POLYTECHNIC UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The presence of ComQ is essential for the activation of the B. subtilis quorum sensing system, but whether this enzyme and its mutants affect vitamin K 2 Metabolic synthesis and resistance to foreign antibiotics are unknown

Method used

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  • A kind of isoprenoid transferase comq mutant, gene, vector, engineering bacteria and preparation method and application
  • A kind of isoprenoid transferase comq mutant, gene, vector, engineering bacteria and preparation method and application
  • A kind of isoprenoid transferase comq mutant, gene, vector, engineering bacteria and preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] B. Subtilis △ COMQ strain construction

[0063] B. Subtilis 168 (Bacillus Subtilis 168) was cultured to an index growth medium, taking 4 ml of bacterial liquid, 10,000 × g centrifugation for 5 minutes to collect the bacteria, discard the medium, and the whole genome DNA was extracted according to the kit. E. coli JM109 containing P7C6 plasmid was cultured to an index growth medium, and 4 ml of bacterial liquid, 8,000 × g centrifuge for 2 minutes, and the cells were collected, and the culture group was discarded, and the plasmid DNA was extracted according to the kit. Take the binder 168 whole genome DNA as a template of the left and right homogenous arm; P7C6 plasmid DNA as an intermediate fragment template, and the product is amplified by PCR by PCR.

[0064] Design specific primers are as follows:

[0065] L-COMQ-F: 5'-TGTTTATCAGACTGTTCGGCT

[0066] L-COMQ-R: 5'-aaattgttatccgctctttctccttgatccgggacagaaa

[0067] COMQ-P7C6-F: 5'-cggatcaaggagaaagagcgataacaattcacag

[0068] C...

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Abstract

The present invention provides an isoprenyl transferase-like ComQ mutant, gene, carrier, engineering bacteria, preparation method and application, the amino acid sequence of the mutant is as SEQ ID NO.2, SEQ ID NO.4, SEQ ID Shown in NO.6 or SEQ ID NO.8. The nucleotide sequence is shown as SEQ ID NO.1, SEQ ID NO.3, SEQ ID NO.5 or SEQ ID NO.7. In the mutant provided by the invention, the protein expression level is significantly reduced, and vitamin K 2 and nattokinase biosynthetic ability has been improved.

Description

Technical field [0001] The present invention relates to the field of molecular biology, more particularly to a ComQ isoprenoid transferase mutants, genes, vectors, bacteria and engineering preparation method and application. Background technique [0002] Recent studies have found that there is a signal exchange system between bacteria that quorum sensing plays a vital role in bacterial morphology and functional expression. By monitoring the concentration of bacterial quorum sensing system detects timing surroundings specific signaling molecules and the concentration of the change in the number of homogeneous or heterogeneous bacterial quorum sensing system upon detecting the quantity or concentration around the bacteria reaches a certain value, a signal will be secreted by issuing a corresponding signal molecule to induce expression of specific genes in bacteria, changing the way the coordination between the behavior of bacteria, nutrients and stimulate signal transduction in bac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C12N15/54C12N15/75C12N15/10C12N1/21C12P7/66C12N9/56C12R1/125
CPCC12N9/1085C12N15/75C12N15/1031C12P7/66C12N9/54C12Y304/21062C12Q2531/113
Inventor 刘艳李伟吴静薛正莲王洲宋平胡刘秀周梦洁胡汶松汪剑
Owner ANHUI POLYTECHNIC UNIV
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