摘要
在丙型肝炎病毒(HCV)eDNA克隆的过程中,将eDNA的PCR产物克隆入载体pGEM-3Zf的SmaI酶切位点,使用PCR法标记的Digoxigenin cDNA探针,通过原位杂交筛选特异的重组体。实验证明用此法进行cDNA克隆具有高度的特异性和实用价值,尤其适用于cDNA PCR产物量小的情况。改进的方法比其它方法更为快速、简便。应用该法,我们成功地克隆并表达了HCV基因组的两个区(C和NS3基因)。
During HCV cDNA cloning, cDNA PCR products were cloned into Sma I site of vector pGEM-3Zf,and then specific recombinants were scre-ened by clony hybridization using digoxigenin cDNA probes which were labelled in PCR. Our work demomstrated that this method was highly specific and valuable for the process of cDNA cloning, especially for small amount of cDNA PCR product. Also modified method was more rapid and simpler than others. With this method, we had been successful in the cloning and expression of two regions (C & NS3 genes ) of HCV genome.
出处
《病毒学报》
CAS
CSCD
北大核心
1993年第3期269-273,共5页
Chinese Journal of Virology
关键词
原位杂交
探针
丙型肝炎病毒
In situ hybridization, Degoxigenin-labelled probe, Hepatitis C virus
