摘要
构建HCVla/1b嵌合型全长cDNA克隆,进行体外转录,脂质体法转染HepG2细胞,以RT-PCR法检测HCV正、负链RNA,Western印迹检测HCV蛋白表达。结果表明,细胞在转染后8代(约35d)内,能间断检测到HCV正、负链RNA以及相对分子质量约70000的HCVNS3蛋白,证明该HCV嵌合体可以在细胞中复制和表达。本研究表明含有该嵌合型全长cDNA的质粒可以为后续HCV的研究提供大量可重复的性质均一的病毒模板,有助于深入了解HCV的复制机制。
A chimeric full-length HCV Type 1a/1b cDNA clone was constructed. The chimeric genome encoded the polyprotein of genotype 1a and replicated via 5' and 3' untranslated region of genotype 1b. The RNA transcripts prepared in vitro was transfected into HepG2 cells by lipofectamine. HCV RNA, positive and negative strands and NS3 protein could be detected by RT-PCR and Western blot in transfected cells in about 35 d respectively. These results suggested that transcripts derived from the chimeric HCV cDNA clone were infectious. This cDNA clone containing chimeric full-length HCV Type 1a/1b genome could provide plentiful and homogeneous viral templates for genetic and functional analyses of HCV replication.
出处
《生物技术通讯》
CAS
2005年第2期121-123,共3页
Letters in Biotechnology
基金
国家自然科学基金项目(30371280)
第四军医大学博士课题(2003005)
