论文部分内容阅读
目的 培养和鉴定重组腺病毒载体 Ad- NT- 3.方法 在 2 93细胞中培养扩增 Ad- NT- 3,用组织培养半数感染量法测定其滴度 .从病毒培养上清中提取 DNA,用聚合酶链反应技术扩增 NT- 3部分基因片段 ,将扩增产物克隆并进行序列分析以测定 Ad- NT- 3中 NT- 3的存在及种属 .结果 Ad-NT- 3扩增后获得了较高滴度的病毒 .从形态学证实 Ad- NT-3含有复制能力的腺病毒 ;聚合酶链反应技术证实 Ad- NT- 3含有 NT- 3基因 ;序列分析明确该 NT- 3基因系人的 NT- 3基因 .结论 Ad- NT- 3重组腺病毒含有人神经营养素 - 3基因及有复制能力的腺病毒
Objective To culture and identify Ad-NT-3 recombinant adenovirus vector.Methods Ad-NT-3 was cultured and expanded in 293 cells, and its titer was determined by half infection in tissue culture.DNA was extracted from the virus culture supernatant, The gene fragment of NT-3 was amplified by polymerase chain reaction (PCR), and the amplified product was cloned and sequenced to determine the existence and species of NT-3 in Ad-NT-3.Results After Ad-NT-3 amplification The virus with high titer was obtained. The adenovirus replication-competent Ad-NT-3 was confirmed by morphology. Ad-NT-3 contained NT-3 gene by polymerase chain reaction. The sequence analysis confirmed that the NT-3 gene Human NT-3 gene.Conclusion Ad-NT-3 recombinant adenovirus contains human neurotrophin-3 gene and replication-competent adenovirus