广东麻鸭DHBV全基因克隆及新读码框的发现

选取3份DHBV阳性广东麻鸭血清提取DHBVDNA,设计一对扩增DHBV全基因序列引物Q1和Q2,扩增并克隆DHBV全基因序列,测序并运用相关计算机软件及方法分析获得的全基因序列。结果表明,广东麻鸭DH-BV全长为3027bp。提交GenBank后获得的收录号分别为:AY433937、AY521226、AY521227(来自1号血清);AY392760、AY536371(分别来自2、3号血清)。AY521227的PreS/SORF出现了单碱基突变,在PreC/CORF之前发现一个新的ORF,暂命名为HORF,HORF也同时存在于GenBank中储存的另外8个DHBV全基因序列中。系统发育分析表明,广东麻鸭DHBV在分化程度上是目前储存于GenBank中的DHBV全序列中最高的。成功克隆了广东麻鸭DHBV全基因序列,序列分析为进一步研究广东麻鸭DHBV提供了有益的信息。

Cloning of DHBV Complete Genome from Guangdong Brown Ducks and Finding of a New ORF

3 samples of Duck hepatitis B virus (DHBV) positive serum from Guangdong brown ducks were chosen to be experimental samples. One pair of primers named Q1 and Q2 were designed to amplify the DHBV complete genome. Following the DHBV complete genomes from the 3 DHBV positive serum samples were amplified and cloned, these DHBV complete genomes were sequenced and then analyzed through related software and related molecular biology methods. The results showed that all the 5 complete genomes from Guangdong brown ducks were 3027 nucleotides long. After submitting to the GenBank, the accession Number were attained as: AY433937, AY521226, AY521227(from DHBV positive serum Number 1.);AY392760, AY536371(from DHBV positive serum Number 2 and 3 respectively). One single nucleotide mutation was occurred in the PreS/S ORF of AY521227. A new ORF was found in the upper reaches of PreC/C ORF and named as HORF temporarily. The HORF also exists in the other 8 DHBV complete genomes from Genbank. Phylogenetic analyze indicated that the differentiation degree of the DHBV complete sequences from Guangdong brown ducks were highest in all the DHBV complete sequences exist in the Genbank presently. The successful clone and analysis of DHBV complete genome from Guangdong brown ducks provide further studies with helpful information.