产气荚膜梭菌α-β融合基因的高效表达  

High-Level Expression of Fusion Gene of Closrtidium perfringens of Alpha-Toxin and Beta-Toxin

作　　者：许崇波[1] 赵宝华[2] 卫广森[3] 王卓[3] 王文成[3] 

机构地区：[1]解放军军需大学军事兽医研究所,吉林长春130062 [2]河北师范大学生命科学院,河北石家庄050016 [3]辽宁省益康生物制品厂,辽宁辽阳111000

出　　处：《中国预防兽医学报》2001年第5期356-359,共4页Chinese Journal of Preventive Veterinary Medicine

摘　　要：用PCR从含产气荚膜梭菌α毒素基因的质粒pXETA1中扩增出α毒素基因 ,用NcoI和BamHI双酶切该α毒素基因 ,回收 0 .95kb的α毒素基因片段 ,再用NcoI和BamHI双酶切含产气荚膜梭菌 β毒素基因质粒pXCPAB2 ,与上述回收的α毒素基因片段连接 ,转化至受体菌BL2 1(DE3)中。经NcoI、BamHI、NcoI酶切反应鉴定和核苷酸序列分析证实 ,获得了理想重组质粒pXCPAB2 ,该重组质粒含有α β融合基因。重组菌株BL2 1(DE3) (pXCPAB2 )经IPTG诱导后 ,其表达产物经ELISA检测和SDS_PAGE分析 ,结果表明重组菌株可以高效表达α β融合蛋白 ,该融合蛋白占菌体总蛋白的 2 2 .14%。Alpha_toxin gene was amplified from plasmid pXETA1 Containing Closrtidium perfringens alpha_toxin gene by Polymerase Chain Reaction (PCR),PCR products were cleaved with restriction endonucleases NcoI and BamHI and recovered. The 3 terminus of α toxin gene were genetically fused to the 5 terminus of β toxin gene of Closrtidium perfringens. The recombinant plasmid pXCPAB2 was studied in detail by restriction endonuclease analysis and nucleotide sequencing.The results have shown that the recombinant plasmid carried α β fusion gene.By transformation of BL21(DE3),we got recombinant strain BL21(DE3)(pXCPAB2).The recombinant strain could produce α β fusion protein by ELISA and SDS_PAGE.After recombinant stain was induced by IPTG, its expressed product was about 22.14% of total cellular protein by SDS_PAGE and thin_layer gel scanning analysis.

关 键 词：产气荚膜梭菌 Α毒素 Β毒素 融合基因 基因表达 

分 类 号：S852.61[农业科学—基础兽医学]