摘要
目的建立凋亡素基因克隆及原核表达的方法。方法根据已报道的鸡贫血病毒凋亡素基因设计合成一对引物 ,通过PCR扩增 ,构建重组质粒pUC18 VP3,将其与pET30质粒分别用限制性内切酶NdeⅠ、EcoRⅠ消化 ,构建原核表达载体pET30 VP3,用以转化感受态的E .coliDE3,经IPTG化学诱导表达 ,表达产物进行SDS PAGE电泳鉴定。结果凋亡素基因在E .coliDE3中获得了大量表达。结论该结果对凋亡素进一步开发为抗肿瘤新药具有重要意义。
PurposeTo establish a method for the cloning and expression of apoptin gene.MethodsAccording to the known sequence of apoptin gene of chicken anemia virus(CAV),a pair of primers for amplifying the complete VP3 were designed and synthesized. The fragment of VP3 was amplified by PCR and was jointed into plasmid pUC18. The recombinant vector and plasmid pET30 were digested by Nde Ⅰ and EcoR Ⅰ enzyme. The expression plasmid pET30-VP3 was constructed,then the recombinant vector was transformed in E.coli DE3 and was induced to express in E.coli DE3 by IPTG.ResultsThe expression product was identified by SDS-PAGE.It proved that a great quantity of apoptin was expressed highly in E.coli DE3.ConclusionThe result has the great value to further develop a safe,effective candidate for cancer therapy.
出处
《中国生化药物杂志》
CAS
CSCD
2004年第5期278-281,共4页
Chinese Journal of Biochemical Pharmaceutics
关键词
凋亡素
克隆
表达
抗肿瘤
apoptin
cloning
expression
candidate for cancer therapy
