摘要
目的 :利用限制性显示 (RD)技术对丙型肝炎病毒 1b亚型 (HCV 1b)基因片段进行克隆与分析。方法 :用限制性内切酶Sau3AⅠ消化HCV 1bcDNA ,所得的限制性内切酶片段物进行RD PCR扩增 ,扩增后的产物克隆至 pMD18 T载体并进行快速鉴定。 结果 :得到 2 0个大小均一 (2 0 0～ 10 0 0bp)的限制性片段 ,测序结果表明 ,属于HCV 1b基因。结论 :RD技术能快速收集大量长度适宜、大小相对均一的病毒基因片段 。
Objective: To clone and analyze the HCV-1b gene fragments isolated by Restriction Display technique(RD) Methods:Restriction enzyme Sau3A Ⅰwas chosen to digest the full-length HCV-1b cDNAs. The products were classified and re-amplified by RD-PCR, the RD-PCR fragments were purified and cloned into the pMD18-T vectors. The recombinant plasmids were extracted from positive clones and the target gene fragments were sequenced. Results:The target HCV gene fragment ranging from 200 to 1000bp were isolated and sequenced which proved to be gene fragments derived from the HCV-1b genome. A total of 20 different cDNA fragments were quickly obtained. Conclusion:RD technique is of great value in obtaining a large number of size-comparable gene probes, which are suitable for the construction of cDNA fragments library and preparation of microarray probes.
出处
《军医进修学院学报》
CAS
2004年第5期327-329,共3页
Academic Journal of Pla Postgraduate Medical School
基金
国家自然科学基金资助项目 ( 3 9880 0 3 2 )
广州市重点科技攻关项目 ( 99 Z 0 2 2 0 1)
