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为估计从毛蚶中提取的甲肝病毒的VP_(1)N端核苷酸和氨基酸的变异,本实验用DNA多聚酶链反应特异性地扩增该区域基因,并将其次级克隆到M13载体作双脱氧核苷酸序列分析。与发表的HM-175、CR326和HAS-15株相应区域比较,除HAS-15株从第76位至93位存在18个核苷酸缺失外,测得的204个核苷酸序列中分别存在15、12和4个碱基替换。其中大多数碱基替换发生在不引起氨基酸改变的第三密码子上,仅第86、91和176位核苷酸改变在第一或第二密码子上。据此推导出不同地区甲肝病毒株的VP_(1)N端前68个氨基酸,除HAS-15株有6个氨基酸缺失外,只存在1至2个氨基酸区别。考虑到病毒是否经传代培养,无明显证据表明绒猴传代与这一区域核苷酸变异有关。
In order to estimate the variation of the nucleotide and amino acid at the VP_ (1) N terminal of Hepatitis A virus isolated from Raucus, DNA polymerase chain reaction (PCR) was used to amplify the gene of this region and subcloned into M13 vector Deoxynucleotide sequence analysis. Compared with the corresponding regions of the published strains HM-175, CR326 and HAS-15, 204 nucleotide sequences were found to be present separately except for the 18 nucleotide deletions from positions 76 to 93 of the HAS-15 strain 15, 12 and 4 bases. Wherein most of the base substitutions occur on the third codon that does not cause an amino acid change, only the 86th, 91st, and 176th nucleotides change on the first or second codon. According to this, the first 68 amino acids of the VP_ (1) N-terminal of hepatitis A virus strains in different regions were deduced. There were only 1 to 2 amino acid differences except that there were 6 amino acid deletions in HAS-15. Considering whether the virus is subcultured, there is no obvious evidence that the passage of marmosets is related to the nucleotide variation in this region.