摘要
目的构建并鉴定pAd/CMV/V5-DEST-p16重组腺病毒质粒。方法合成人p16-INK4表达基因,构建pENTR1A-p16质粒。通过LR反应,入口克隆pENTR1A-p16质粒的外源性合成的修饰后的p16cDNA,取代目的质粒pAd/CMV/V5-DEST中的ccdB基因,形成表达克隆pAd/CMV/V5-DEST-p16,测序证实质粒含有目的基因。结果构建了p16重组腺病毒质粒,为研究p16-INK4的作用创造了条件。结论用通路克隆系统构建重组p16腺病毒质粒稳定、可靠、方便。
Objective To construct recombinant p16 Adenovirus plasmid. Methods The sequence of the p16 INK4 suppressor gene from human was artificially synthesized. And pENTR 1A-p16 plasmid, the entry clones,were generated by serial tests. Along with the LR recombination reactions finished and the ccdB gene in the pAd/ CMV/V5-DEST plasmid replaced by the p16-INK4, the pAd/CMV/V5-DEST-p16 plasmid, the expression clones, were created. The sequencing of the artificial synthesis sequence was right. Results The construction of the recombinant p16 Adenovirus plasmid was successful which provided a basis for the study of the function of the suppressor gene p16-INK4. Conclusion It is stable, reliable and convenient to construct the recombinant p16 Adenovirus vector with gateway cloning system.
出处
《安徽医科大学学报》
CAS
北大核心
2006年第3期258-261,共4页
Acta Universitatis Medicinalis Anhui
基金
安徽省自然基金资助课题(编号:000-44322)
