Human defensins mHD-5 gene and related application thereof

A human defensin and gene technology, applied in the field of genetic engineering, can solve the problems of cumbersome procedures, expensive chemical synthesis rules, and high cost, and achieve the effects of simple culture and subsequent operation processes, reduced human toxicity and side effects, and easy portability.

Inactive Publication Date: 2017-02-22
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, although defensins play a certain role in relieving intestinal inflammation and synergistically regulating immunity, the extraction of defensins is mainly obtained from natural resources, which restricts its wide application due to high cost, low yield, and cumbersome procedures. Synthetic rules are relatively expensive and difficult to apply clinically
Using molecular biology techniques, expressing defensins in expression systems such as Escherichia coli and yeast is considered a feasible strategy, but its safety and subsequent purification of antimicrobial peptides limit the practical application of these expression systems

Method used

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  • Human defensins mHD-5 gene and related application thereof
  • Human defensins mHD-5 gene and related application thereof
  • Human defensins mHD-5 gene and related application thereof

Examples

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Embodiment 1

[0031] Example 1 Optimized human defensin 5 (mHD-5) gene and Lactococcus lactis expression plasmid construction

[0032] 1. Gene optimization of human defensin 5

[0033] Referring to the natural gene sequence of human defensin 5 (HD-5) from the defensin family (GeneBank: NM_021010.1), based on the characteristics of the Lactococcus lactis genome, it was aligned according to the GC content and rare codon usage. Gene optimization was carried out to obtain the optimized gene sequence mHD-5 (SEQ ID No: 1), the amino acid sequence of which is shown in figure 1 .

[0034] 2. Construction of Lactococcus lactis expression plasmid NZ9000-SHD5

[0035] see figure 2In order to construct the experimental process of recombinant Lactococcus lactis NZ9000-SHD5, the optimized gene mHD-5 was cloned into the plasmid pMD19T-simple (Sangon Bioengineering (Shanghai) Co., Ltd.) to obtain the pMD19TS-mHD-5 plasmid, which was used Transformed into Escherichia coli Top10 by chemical transformati...

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Abstract

The invention discloses a human defensins mHD-5 gene and related application thereof. The human defensins mHD-5 gene has a base sequence as shown in SEQ ID NO:1. Recombinant lactococcus lactis comprises the optimized human defensins mHD-5 gene. A Western Blot result shows that the recombinant lactococcus lactis NZ9000-SHD5 disclosed by the invention can implement secretory expression of the recombinant defensins mHD-5 gene; meanwhile, an in-vitro experiment result shows that the recombinant defensins mHD-5 gene subjected to secretory expression by the recombinant lactococcus lactis NZ9000-SHD5 has an effect of effectively alleviating inflammation; compared with a conventional treatment mode, the lactococcus lactis recombinantly expressing the defensins gene combines the advantages of probiotics and defensins, and is creatively applied to reduce disease activity and alleviate inflammation of the inflammatory bowel disease.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and more specifically, the invention relates to an optimized human defensin mHD-5 gene and related applications. Background technique [0002] Inflammatory bowel disease (Inflammatory bowel disease, IBD) is a non-specific chronic intestinal inflammation of unknown etiology, including ulcerative colitis (Ulcerative colitis) and Crohn's disease (Crohn's disease), its etiology and pathogenesis are still unknown Fully clarified, it has become a common disease of the digestive system in recent years. Although the pathogenesis of IBD is not yet clear, relevant data prove that the initiation and recurrence of inflammation are related to the intestinal immune response, and the expression of TNF-α and IFN-γ in the intestine increases. At the same time, highly expressed inflammatory factors in intestinal inflammation are also potential pathogenic factors, which damage the mucosal barrier funct...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/74C12N15/64C12N1/21A61K38/17A61P1/00
CPCC07K14/4723A61K38/00C12N15/746C12N2800/101
Inventor 陈烨陈卫谭嘉圣曾丽珊
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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