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Cultivated silkworm peptidoglycan identification protein gene

A technology for identifying proteins and peptidoglycans, applied in the field of protein genes, can solve the problems that there are no research reports on the peptidoglycan recognition protein genes of silkworms

Inactive Publication Date: 2008-03-12
SOUTHWEST UNIVERSITY
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AI Technical Summary

Problems solved by technology

So far, there is no research report on the peptidoglycan recognition protein gene of silkworm

Method used

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Experimental program
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Embodiment

[0040]The silkworm genome database and EST database were predicted to obtain the full-length ORF of the PGRPs gene, and primers were designed accordingly. The upstream primer sequence: 5'CGCG GAT CCT AGT CCA ATA AAA TTA GTT AG 3'downstream primer sequence: 5'CCCAAG CTT ATC ATC CCA ATT CAG TGA AA 3'; the five-instar and three-day-old silkworm Dazao, after being induced by PGN for 12 hours, the fat body was taken, frozen and stored in liquid nitrogen; RNA was extracted by RNAiso reagent (TaKaRa), and cDNA was synthesized; amplified by PCR The BmPGRPs gene fragment was obtained, the amplified fragment was cloned into PMD18-T Simple Vector, and then the sequence was verified; the similarity comparison was completed on the NCBI site with BLAST, and the BLAST analysis showed that the PGRPs gene was relatively conservative, which had not been reported in related research on silkworms , is a new gene; the full-length ORF fragment of the PGRPs gene was cloned into the prokaryotic expres...

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Abstract

The invention provides an identifying protein gene for a silkworm peptidoglycan. First of all, predicating PGRPs gene by using a silkworm gene database and EST database source, then extracting PGA-induced 5-instar silkworm epiploon, synthesizing CDNA, and cloning gene. PGRPs gene can obviously kill bacteria, and PGRPs pertain solution of low concentration can also suppress bacteria growth. The congenital immunity identifying receptor of silkworm activates the congenital immunity and adjusts the antibiotic peptide expression, and can be used for culturing antibiotic silkworms.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein gene derived from silkworm innate immune recognition receptors, capable of recognizing bacterial cell wall peptidoglycan, activating the silkworm immune system and antibacterial. Background technique [0002] my country started silkworm breeding as early as 5,000 years ago and is the birthplace of sericulture. With the continuous development of bioengineering technology, the comprehensive utilization level of silkworm will be improved. The components of microbial cell walls are highly efficient activating molecules of innate immunity, such as peptidoglycan (PGN) of Gram-positive bacteria. PGN is a polymer composed of an amino sugar backbone and a peptide chain, and its structural units are composed of β-(1,4)-linked N-acetylglucosamine and N-acetylmuramic acid disaccharide units and tetrapeptide subunits and peptide bridge composition. The difference in the composition a...

Claims

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Application Information

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IPC IPC(8): C12N15/12C07K14/435
Inventor 刘春程廷才夏庆友向仲怀
Owner SOUTHWEST UNIVERSITY
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