用PCA方法选择HBV DNA多聚酶TP区VH抗体
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于俊岩, 兰 林, 王宇明
乙型肝炎病毒; 末端蛋白; VH抗体; PCA方法,用PCA方法选择HBVDNA多聚酶TP区VH抗体,于俊岩, 兰林, 王宇明,于俊岩
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于俊岩, 兰林, 王宇明, 第三军医大学西南医院感染病研究所 重庆市 400038
于俊岩, 男, 1969-07-17生, 山西屯留县人, 汉族, 1992年长治医学院毕业, 2001年山西医科大学传染病学硕士研究生毕业, 2003年至今在第三军医大学攻读传染病学博士学位, 讲师, 主要从事慢性乙型肝炎发病机理研究.
国家自然科学基金资助项目, No.30100159
通讯作者: 兰林, 400038, 重庆第三军医大学西南医院感染病研究所. Llin6624@hotmail.com
电话: 023-68752193
收稿日期: 2005-04-21 接受日期: 2005-06-01
Isolation of the variable fragments of heavy chain against the terminal protein region of hepatitis B virus polymerase with protein fragment complementation assay
Jun-Yan Yu, Lin Lan, Yu-Ming Wang
Jun-Yan Yu, Lin Lan, Yu-Ming Wang, Department of Infectious Diseases, Southwest Hospital, the Third Military Medical University, Chongqing 400038, China
Supported by National Natural Science Foundation of China, No. 30100159
Correspondence to: Dr. Lin Lan, Department of Infectious Diseases, Southwest Hospital, the Third Military Medical University, Chongqing 400038, China. Llin6624@hotmail.com
Received: 2005-04-21 Accepted: 2005-06-01
Abstract
AIM: To isolate the variable fragments of heavy chain (VH) against the terminal protein (TP) region of hepatitis B virus (HBV) polymerase (Pol) with protein fragment complementation assay (PCA).
METHODS: The plasmid pPNL6, which contained scFv antibody library, was extracted from Yeast Display scFv Antibody Library to amplify the VH antibody fragment. The HBV DNA secreted by HepG2.2.15 cells was used as the template to amplify HBV DNA TP. Then the VH antibodies were selected with PCA using the TP region as the antigen.
RESULTS: After amplification, TP region with a length of 540 bp were obtained. Three TP antigen-specific VH antibodies were amplified by PCA, and then they were translated into amino acids according to the DNA sequences and Werner Müller databank ......
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