机构地区: 宁夏大学生命科学学院
出 处: 《生物技术》 2003年第4期1-2,共2页
摘 要: 利用聚合酶链式反应 (PCR)技术 ,从A型产气荚膜梭菌染色体基因组中扩增了 1.2kb的α毒素基因。通过T4 DNA连接酶 ,将纯化的PCR产物与载体pGEM -T连接 ,转化至受体菌JM10 9中。经NcoI EcoRI和BamHI EcoRI双酶切分析 ,证明重组质粒pXCPA0 2中含有A型产气荚膜梭菌α毒素全基因。经核苷酸序列分析 ,明确了克隆的α毒素基因在重组质粒中的连接向位且核苷酸序列是正确的。 Alpha-toxin gene was amplified from genomic DNA of Clostridium perfringens type A by polymerase chain reaction(PCR).PCR product was inserted into vector pGEM-T directively.The recombinant plasmid pXCPA02 was identified by restriction endonucleases Nco I/ Eco RI and Bam HI/ Eco RI analysis and nucleotide sequencing.The results showed that alpha-toxin gene fragment of 1.2kb has been cloned with a positive nucleotide sequence.