Hu Ji-li, Tang Ni, Huang Ai-long
Institute of Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China.
Zhonghua Gan Zang Bing Za Zhi. 2003 Jun;11(6):341-3.
To clone and analyze duck hepatitis B virus genome from Chongqing brown duck.
Duck hepatitis B virus (DHBV) DNA extracted from a Chongqing brown duck was amplified by PCR and cloned into PGEM-T vector using T-A clone method. The sequence of this DHBV genome was analyzed with some softwares after identified.
The duck hepatitis B virus genome from Chongqing brown duck (DHBVcq), which was 3 024 nucleotides long, contained three ORFs whose onset and end nucleotides were in accord with those of HPUGA, encoding P, PreC/C and PreS/S protein respectively. Comparison of this strain with other DHBV reported in GenBank showed that the homology of DHBVcq and M32990 got the highest score of 94.9% at nucleotide level, while DHBVcq and DHBVCG got the least (89.8%). Most of the conserved regulation nucleotides and amino acids sequence found in other DHBV were also identified in DHBVcq. The epsilon region of DHBVcq, which was important for encapsidation of pgRNA and synthesis of minus-strand DNA, differed from that of most other DHBV strains, forming a stem-loop conformation with a three- nucleotides upper stem rather than a common nine-nucleotides one in free status.
The successful clone and analysis of DHBVcq provide further studies with helpful information.
克隆并分析重庆麻鸭的鸭乙型肝炎病毒基因组。
从重庆麻鸭中提取鸭乙型肝炎病毒(DHBV)DNA,通过PCR扩增,采用T-A克隆法将其克隆至PGEM-T载体。鉴定后,使用一些软件对该DHBV基因组序列进行分析。
重庆麻鸭的鸭乙型肝炎病毒基因组(DHBVcq)长3024个核苷酸,包含3个开放阅读框,其起始和终止核苷酸与HPUGA一致,分别编码P、PreC/C和PreS/S蛋白。该毒株与GenBank中报道的其他DHBV比较,结果显示DHBVcq与M32990在核苷酸水平上的同源性最高,为94.9%,而与DHBVCG的同源性最低(89.8%)。在其他DHBV中发现的大多数保守调控核苷酸和氨基酸序列在DHBVcq中也有发现。DHBVcq的ε区对pgRNA的包装和负链DNA的合成很重要,它与大多数其他DHBV毒株的ε区不同,形成了一个茎环结构,其上游茎为三个核苷酸,而不是游离状态下常见的九个核苷酸。
DHBVcq的成功克隆和分析为进一步研究提供了有用信息。