摘要
将分别携带SHIV-KB9 (SIV/HIV-1 KB9) 基因组的3′端和5′端的两个半长克隆,体外连接成SHIV-KB9全基因组克隆。含有全长基因的质粒培养时易发生同源重组和缺失,采用JM109作为宿主菌以及30℃、低转速的培养条件,可保持质粒的稳定性。通过PCR , RT-PCR 和猴免疫缺陷病毒(SIV) gag p27 核心抗原滴度检测表明:感染性克隆SHIV-KB9可有效在人、恒河猴及食蟹猴的外周血单核细胞中复制。
SHIV-KB9 chimeric virus was constructed by two constructs representing the 3?and 5?fragments of the viral genome. This full-genome SHIV-KB9, with large size and long terminal repeat sequence (LTR), could be maintained stable in E.coli JM109 strain in the cultivation condition of 30℃ and 180r/min. The infectivity of the full-length SHIV-KB9 in human peripheral blood mononuclear cells (PBMC), cynomolgus and rhesus monkey PBMC was characterized by polymerase chain reaction (PCR), reverse transcription PCR (RT-PCR) and Simian immunodeficiency virus (SIV) p27 gag antigen titration assay.
出处
《中国病毒学》
CSCD
2004年第2期114-119,共6页
Virologica Sinica
基金
美国国立健康研究所CIPRA资助(U19 AI051915)~~
