产气荚膜梭菌α-β融合基因的构建  

Construction of Fusion Gene of Clostridium perfringens of Alpha Toxin and Beta Toxin

作　　者：许崇波[1] 赵宝华[2] 卫广森[3] 蒋玉文[4] 王卓[3] 

机构地区：[1]解放军军需大学军事兽医研究所 [2]河北师范大学生命科学院 [3]辽宁省益康生物制品厂 [4]中国兽药监察所北京100080

出　　处：《中国兽医学报》2001年第4期341-343,共3页Chinese Journal of Veterinary Science

基　　金：辽宁省自然科学基金资助项目

摘　　要：用 PCR从含产气荚膜梭菌β毒素基因的质粒 p XETB2中扩增出β毒素基因 ,Nco 和 Bam H 双酶切该 β毒素基因 ,回收 0 .93kb的 β毒素基因片段 ,再用 Nco 和 Bam H 双酶切含产气荚膜梭菌 α毒素基因质粒 p XETA1 ,与上述回收的 β毒素基因片段连接 ,转化至受体菌 BL2 1 ( DE3)中。经 Nco 、Bam H 、Not 酶切反应鉴定和核苷酸序列分析证实 ,获得的重组质粒 p XCPAB1含有 α-β融合基因。重组菌株 BL2 1( DE3) ( p XCPAB1 )表达产物经 ELISA检测和 SDS-PAGE分析 。Beta toxin gene was amplified from plasmid pXETB2 containing Clostridium perfringens beta toxin gene by polymerase chain reaction (PCR), PCR products were cleaved with restriction endonucleases NcoⅠ and BamHⅠ and recovered. The 3′ terminus of β toxin gene was genetically fused to the 5′ terminus of α toxin gene of Clostridium perfringens. The recombinant plasmid pXCPAB1 was characterized by restriction endonuclease analysis and nucleotide sequencing. The results showed that the recombinant plasmid bore the α β fusion gene. By transformation of BL21(DE3), we got recombinant strain BL21(DE3)(pXCPAB1). The recombinant strain was proved to be able to produce α β fusion protein by ELISA and SDS PAGE.

关 键 词：产气荚膜梭菌 Α毒素基因 β毒素基因 融合基因 基因克隆 动物 坏死性肠炎 

分 类 号：S858.64[农业科学—临床兽医学] S855.12[农业科学—兽医学]