
Cloning and sequence analysis of full-length cDNA of hepatitis D virus in China
MA Jian1, GU Wen-Jie1, JIA Xue-Rong2, HUANG Wei-Jin1, LIANG Zheng-Lun1, WANG You-Chun1
Journal of Microbes and Infections ›› 2015, Vol. 10 ›› Issue (6) : 345-350.
Cloning and sequence analysis of full-length cDNA of hepatitis D virus in China
The present paper aims to clone and analyze the sequence of full-length cDNA of hepatitis D virus (HDV) in China. HDV RNA was extracted from three HDV seropositive samples. cDNA was gotten by reverse transcription-polymerase chain reaction (RT-PCR). Four overlapped fragments were amplified by nested PCR, and the products were sequenced. The full-length sequences were joined by DNAStar. The full-length cDNA of HDV was amplified by overlapping PCR using designed primers and connected to the cloning T vector. Three strains of HDV were successfully amplified and cloned. The three HDV strains were all genotype I. The nucleic acid sequence homology of the three HDV strains was over 98%. The nucleic acid molecules of three strains had at least 84% sequence homology to genotype Ib. But they had over 98% sequence homology to genotype I Chinese strain X77627. They had no more than 77% and 65% sequence homology compared with genotype II and genotype III respectively. In conclusion, we cloned full-length cDNA of three strains of HDV, which could be helpful for the further study of HDV molecular biology.
Hepatitis D virus / Polymerase chain reaction / Full-length genome
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