摘要
目的 克隆人腺病毒 3型 (Ad3)六邻体 - L 1(Hexon- L 1)、六邻体 - L 2 (Hexon- L 2 )区基因 ,构建含有该基因的重组质粒 ,并进行测序鉴定 ,为进一步构建表达载体 ,制备基因工程疫苗打下基础。方法 从 Ad3感染的 He L a细胞中提取 Ad3DNA,用 PCR方法扩增 Hexon- L 1、Hexon- L 2基因 ,将得到的片段定向克隆到克隆载体 p U C19质粒中 ,对克隆片段进行 DNA测序鉴定。结果 构建了重组质粒 p UC19Hexon,测序结果与 Genebank中该序列进行同源性比较证明克隆的片段为 Ad3六邻体序列。结论 成功克隆了 Ad3的 Hexon- L 1、Hexon- L 2区基因。
Objective To clone the Hexon-L1、Hexon-L2 gene of human adenovirus type 3 for constructing the expression recombinant plasmid and producing genetic engineering vaccine to prevent the adenovirus infection.Methods Ad3 DNA was isolated and purifyied from the cultured HeLa cells.The Hexon fragment was amplified by polymerase chain reaction (PCR) method,and then cloned into vector pUC19 plasmid and charactered by restriction enzyme digestion and sequencing.Results The recombinant pUC19Hexon was constructed. Meanwhile,it was demonstrated that the cloned fragement was Ad3 Hexon sequence indeed by aligning with the sequence of Ad3 in genebank.Conclusions The Ad3 Hexon-L1, Hexon-L2 gene are cloned successfully.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2003年第3期245-247,共3页
Chinese Jouranl of Endemiology
基金
中国医学科学院青年基金资助项目 (93 3 10 14 )
