WU Ying-yun, HONG Kun-xue, CHEN Pei, CHEN Qi-ming, GENG Yun-qi and SHAO Yi-ming. Generation of Full-genome SHIV-KB9 Clone and Its Replication in Human and Monkey Peripheral Blood Mononuclear Cells*[J]. Virologica Sinica, 2004, 19(2): 114-119.
Citation:
WU Ying-yun, HONG Kun-xue, CHEN Pei, CHEN Qi-ming, GENG Yun-qi, SHAO Yi-ming.
Generation of Full-genome SHIV-KB9 Clone and Its Replication in Human and Monkey Peripheral Blood Mononuclear Cells* .VIROLOGICA SINICA, 2004, 19(2)
: 114-119.
全基因组SHIV-KB9克隆的构建及其在人、猴 外周血单核细胞中的复制*
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吴颖运
1,2
,
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洪坤学
2
,
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陈启明
2
,
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耿运琪
1
,
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邵一鸣
1
,
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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。
Generation of Full-genome SHIV-KB9 Clone and Its Replication in Human and Monkey Peripheral Blood Mononuclear Cells*
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1.
1. College of Life Sciences, Nankai University, Tianjin 300071, China
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2.
National Center for AIDS/STD Control and Prevention, Beijing 100050, China
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Corresponding author:
SHAO Yi-ming,
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Abstract
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.
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References
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