Operators, their vectors and their applications

A promoter and promoter sequence technology, applied in the field of bioengineering, can solve the problems of few genetic tools, few types, limited expression activity of promoters, etc.

Active Publication Date: 2022-05-20
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] At present, there are problems such as low protein expression efficiency and few available genetic tools in the existing technology, and the development of high-efficiency promoters that can be used for foreign protein expression is an important way to solve these problems
Inducible promoters commonly used in recent years include tac promoter (Ptac), arabinose promoter, maltose-inducible promoter, propionate-inducible promoter, temperature-sensitive promoter, etc., but the expression activity of these promoters is very limited and can be Less variety to choose from

Method used

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  • Operators, their vectors and their applications
  • Operators, their vectors and their applications
  • Operators, their vectors and their applications

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Embodiment approach

[0055] As one of the embodiments of the present invention: select the gene SEQ ID NO: 6 gene coding sequence ATG upstream 355bp to downstream 62bp sequence region of C. glutamicum BZH 001 to design promoter cloning primers 355ZDA-F and ZDA-R (SEQ ID NO: The 6 gene coding sequence ATG starts at the 501bp base of the SEQ ID NO: 6 sequence, and the SEQ ID NO: 6 gene coding sequence is the 501bp ~ 1355bp of the sequence table SEQ ID NO: 6), the clone promoter sequence and its The 62bp sequence (including ATG) after the downstream gene ATG, and a 15bp RBS sequence (AAAGGAGGACAACTA) and two reverse BsaI restriction sites and a BamH I restriction site were sequentially introduced at the 5' end of the downstream primer ZDA-R , and the BamH I restriction site overlaps with the second Bsa I restriction site cut;

[0056] ZDA-F TTCG AAGCTTC TTTGAAGCGGTGTGTGAC (the underline is the enzyme cutting site Hind III)

[0057] ZDA-R

[0058] CCGG GGATCCGAGACC AAAAAAATTTTTT GGTCTC ACATTAG...

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Abstract

The invention discloses an operon, its carrier and its application, wherein the operon sequence at least includes, the 3' end is located at any base in the 201-257 bp upstream of the ATG coding sequence of the Corynebacterium glutamicum SEQ ID NO: 6 gene The 5' end is located at any base in the upstream 308bp-345bp of the coding sequence ATG of the Corynebacterium glutamicum SEQ ID NO: 6 gene. After the operon of the present invention is induced by the inducer, the protein expression activity is significantly higher than that of the traditional Ptac, and it is an ultra-efficient operon. The operon of the present invention can be used for high-efficiency expression of prokaryote endogenous or exogenous proteins.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and specifically relates to an operon, its carrier and its application. Background technique [0002] Promoter is a key DNA sequence that controls the initiation of gene transcription and plays an important role in regulating gene expression. The promoter mainly controls the transcription level of genes. A large number of studies have proved that in various sequence regions that control gene expression Among them, the influence of the promoter region exceeds 45%, and the operator regulates the expression of the entire promoter. Therefore, it is of great significance to screen highly active operons for the study of gene expression. [0003] At present, there are problems such as low protein expression efficiency and few available genetic tools in the existing technology, and the development of high-efficiency promoters that can be used for exogenous protein expression is an important way t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/77
CPCC12N15/113C12N15/77C07K14/34
Inventor 刘秀霞董贵彬曹煜余心宇邓智威赵子豪杨艳坤白仲虎
Owner JIANGNAN UNIV
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