摘要
目的 :表达志贺毒素 2 A-黄体生成激素释放激素 ( Stx2 A- LHRH)重组毒素 ,探讨其是否具有特异杀伤癌细胞的可能性。方法 :用 PCR方法扩增带有 Nco 和 Eco R 酶切位点的 Stx2 A-LHRH DNA基因 ,克隆至 p ET- 2 0 b( + )质粒中 ,转化大肠杆菌 BL2 1 ( DE3) ,挑取单菌落培养 ,提取质粒、酶切。结果 :经鉴定、测序 ,获得重组质粒 p ET2 0 - SL。结论 :利用分子生物学技术成功地构建了带有 Stx2 A- LHRH重组毒素基因的质粒 ,使其表达 Stx2 A- LHRH重组毒素成为可能 ,为进一步研究导向药物奠定了基础。
Objective:To study the expression and possibility of cytotoxic activity of recombinant toxin Stx2A LHRH in human carcinoma cell. Methods:Stx2A LHRH sequences in which the restriction endonuleases NcoⅠ and EcoRⅠ site was added at the 5′ and 3′ends were amplified by PCR and digested with appropriate restriction enzymes.The digested fragment was subcloned into the vector obtained by digestion of plasmid pET 20b(+) with NcoⅠ and EcoRⅠ. E.coli BL21(DE3) cells were transformed with the recombinant plasmids and cultured in LB medium containing ampicillin, then the plasmid was extracted and digested. Results:Recombinant plasmid pET20 SL was constructed successfully and the clones expressing pET20 SL stablely were identified by digestion and sequencing. Conclusion:In this study,construction of recombinant plasmid encoding chimeric toxin Stx2A LHRH and its expression were completed successfully by molecular biology technique.The finding could open up new vistas in the study on targeted drugs.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2003年第2期177-179,共3页
Journal of Jilin University：Medicine Edition
关键词
志贺毒素2A
黄体生成激素释放激素
重组毒素
质粒
Shiga toxin 2A (Stx2A)
Luteinizing hormore releasing hormone (LH RH)
Chimeric toxin
Plasmid
