源于草鱼肠道抗菌肽的重组表达、分离纯化和抗菌活性分析
Expression, Prufication and Antimicrobial Analysis of Antimicrobial Pepetide Originated from intestine of Grass Carp
本研究根据草鱼肠道中分离得到的抗菌肽氨基酸测序结果，人工合成草鱼抗菌肽基因，并将该合成基因克隆至pET32a载体中，构建成含有含6×His标签的重组质粒pET-AP。重组质粒转化至大肠杆菌BL21中，经IPTG诱导后，进行SDS-PAGE分析。测序结果表明，插入片段的序列和位置均正确。通过Ni2+-NTA柱亲和层析纯化，用肠激酶将纯化的重组蛋白中的His标签切除后回收重组抗菌肽，采用滤纸片方法测定其抗菌活性。结果表明重组抗菌肽能明显抑制大肠杆菌的生长。
ccording to the amino acid sequence of intestine of grass carp, antimicrobial peptide (AP) coding genes were synthesized artificially and cloned into vector pET32a to construct a recombinant plasmid termed pET32AP which contains a 6×His tag. Plasmid pET-AP was transferred to Escherichia. coli strain BL21. SDS-PAGE analysis of recombinant protein was carried out after induced by IPTG. Sequencing results indicated that the sequence and location of insert fragment in pET-AP were correct. Recombination protein was purified by Ni2+-NTA column affinity chromatography. And enterokinase was used to recover the recombinant antimicrobial peptide (AP). Antimicrobial activity assay showed that recombinant AP could inhibit the growth of E. coli.
苏建明、孙科军、史玉婷、陈韬、陶虚谷
生物工程学分子生物学生物化学
草抗菌肽重组表达纯化抗菌活性
ntimicrobial peptideRecombinant expressionPurificationAntimicrobial activity
苏建明,孙科军,史玉婷,陈韬,陶虚谷.源于草鱼肠道抗菌肽的重组表达、分离纯化和抗菌活性分析[EB/OL].(2012-03-13)[2025-08-16].http://www.paper.edu.cn/releasepaper/content/201203-422.点此复制
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