CAJ | 학술논문

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.
장분별휴대SHIV-KB9 (SIV/HIV-1 KB9) 기인조적3′단화5′단적량개반장극륭,체외련접성SHIV-KB9전기인조극륭.함유전장기인적질립배양시역발생동원중조화결실,채용JM109작위숙주균이급30℃、저전속적배양조건,가보지질립적은정성.통과PCR , RT-PCR 화후면역결함병독(SIV) gag p27 핵심항원적도검측표명:감염성극륭SHIV-KB9가유효재인、항하후급식해후적외주혈단핵세포중복제.
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.