期刊信息
- 刊名： 河北师范大学学报（自然科学版）Journal of Hebei Normal University (Natural Science)
- 主办： 河北师范大学
- ISSN： 1000-5854
- CN： 13-1061/N
- 中国科技核心期刊
- 中国期刊方阵入选期刊
- 中国高校优秀科技期刊
- 华北优秀期刊
- 河北省优秀科技期刊
Cloning and Expression of Hemolysin Gene of Human Aeromonas sobira
摘要/Abstract
根据GenBank报道的人源温和气单胞菌溶血素(hly)基因序列设计引物,经PCR扩增,得到大小约为1470 bp的阳性产物.利用BamHⅠ,HindⅢ位点将hly基因片段克隆到pMD19-T载体,构建得到重组质粒pWT-hly,并比较所克隆基因序列的同源性.然后将该基因克隆到原核表达载体pET-28a,构建原核重组表达质粒pWET-hly,转化BL21(DE3)并进行SDS-PAGE分析.结果表明:克隆的hly基因与GenBank报道的溶血素基因同源性为96.19%;重组菌株经IPTG诱导后,hly基因得到了高效表达.
According to the published hemolysin nucleotide sequence from human Aeromonas sobira in Gen-Bank,a pair of primers were designed and synthesized,which were localized in conservative regions of hemolysin gene and were modified with Bam H Ⅰand Hind Ⅲenzyme sizes.A 1470 bp gene fragment was amplified via PCR and the recombinant plasmid pWT-hly was const ructed.Nucleotide sequence analysis indicated that the gene was 1467 bp in length and homology of nucleic acids between this gene and other similar gene in GenBank was 96.19%.The recombinant expression plasmid pWET-hly was const ructed and transformed into E.coli BL-1(DE3).Then the recombinant strain BL-1(DE3)(pWET-hly) was obtained and SDS-PAGE analysis in-dicated that the hly gene was highly expressed when the recombinant strains was induced by IPTG.