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Recombinant plectasin as well as preparation method and application of recombinant plectasin

A technology of recombinant bacteria and recombinant vectors, applied in the field of genetic engineering, can solve the problems of low expression level, difficult to realize industrialized production and the like, and achieve the effects of high antibacterial activity, simple preparation method and high yield

Active Publication Date: 2013-01-30
桂林风鹏生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In the prior art, researchers used different genetic engineering methods to express recombinant plectasin, but the expression level was low, making it difficult to realize industrial production

Method used

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  • Recombinant plectasin as well as preparation method and application of recombinant plectasin
  • Recombinant plectasin as well as preparation method and application of recombinant plectasin
  • Recombinant plectasin as well as preparation method and application of recombinant plectasin

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Embodiment 1

[0031] The preparation of embodiment 1 recombinant antimicrobial peptide of the present invention

[0032] 1. Construction of Recombinant Antimicrobial Peptide Genetic Engineering Bacteria

[0033] 1. Cloning of the target gene fragment

[0034] (1) Extraction of total RNA from P. nigrella

[0035] RNAiso Plus total RNA extraction kit (D9108A) from Takara Company was used.

[0036] (2) Use RT-PCR and site-directed mutagenesis to amplify the plectasin coding sequence. The primers (the underline is the restriction site, the italic part is the mutated base) and the reaction conditions are as follows:

[0037] Forward primer (F):

[0038] 5'- GAATTC GGATCCATTGAAGGCCGCGGCTTTGGCTGCAATGGC-3' reverse primer (R):

[0039] 5'-AGCGGCCGC GTC GAC TTAATAGCATTTGCACACAAAGCCGCCTTTCGCGCAATAGCC-3’

[0040] The reaction conditions are:

[0041]

[0042] (3) DNA fragment recovery: cut out the gel strip containing the target gene, and recover it with a small column gel recovery kit;

...

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Abstract

The invention discloses a recombinant antibacterial peptide which has an amino acid sequence shown in SEQ ID NO.1. The invention also discloses a nucleotide sequence for encoding the recombinant antibacterial peptide, a recombinant vector and recombinant bacteria comprising the nucleotide sequence, and a preparation method and antibacterial application of the recombinant antibacterial peptide. The recombinant antibacterial peptide has high antibacterial activity, and the method for preparing the recombinant antibacterial peptide is simple and high in yield and has good application prospect.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a recombinant plectasin and its preparation method and application. Background technique [0002] In recent years, due to economic and profit-driven, the abuse of antibiotics and the excessive use of antibiotics are common. With the widespread and long-term use of antibiotics, their negative effects have become increasingly prominent. Bacterial drug resistance and drug residues are a serious threat to human health, and it is imperative to seek alternatives to antibiotics. my country has developed green additives such as antimicrobial peptides, prebiotics, saccharoterpenes, oligosaccharides, Chinese herbal medicines, enzyme preparations, acidulants, etc. to replace antibiotics. [0003] Antimicrobial peptides are a kind of non-specific immune response products produced by the host immune system to resist external pathogen infection, and widely exist in insects, plants, animals ...

Claims

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Application Information

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IPC IPC(8): C07K14/37C12N15/31C12N15/63C12N1/21C12P21/02A61K38/16A61P31/04A61P31/10C12R1/645
Inventor 何军陈代文余冰郑萍毛湘冰
Owner 桂林风鹏生物科技有限公司
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