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Virologica Sinica, 32 (2) : 167, 2017
A novel phenotypic assay of hepatitis B virus polymerase with extensive site-specific mutagenesis
1. Institute for Viral Hepatitis, Key Laboratory of Molecular Biology on Infectious Diseases, Ministry of Education, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
2. Department of Microbiology and Immunology, Indiana University School of MedicineIndianapolis 46202, USA
3. Collaborative Innovation Center for diagnosis and treatment of infectious diseases(CCID), Hangzhou 310058, China
 Correspondence: zhangwenlu@cqmu.edu.cn;hujieli1977@163.com
(1984.76KB)  (111.84KB)  
Abstract
In summary, we developed a novel mutagenesis phenotypic assay with the following characteristics: (1) acquisition of extensive site-specific mutations through a PCR step and FSR; (2) production of a lentivirus pool expressing different polymerase mutants via one-time lentivirus packaging; (3) generation of stable cell lines that replicate HBV DNA by integrated polymerase mutants, which can help to establish an experimental system with lower variation and higher convenience. This randomized phenotypic assay of HBV polymerase provides a useful tool for assessing the replication fitness and drug susceptibility of HBV variants and quasi-species, which may offer new insight for the NUC therapy and the prognosis of patients with chronic hepatitis B.
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Received: 24 Apr 2017  Accepted: 24 Apr 2017  Published online: 24 Apr 2017
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