摘要
把一个新鸡贫血病毒(CAV)的vp1基因通过PCR扩增,然后克隆到质粒载体pUC18上.vp1基因包含1 347个碱基对,并且推定的VP1蛋白氨基酸序列含有449个氨基酸.通过DNA BLAST软件把该基因的序列数据与在(Gen-Bank中发表的其他vp1基因比较显示,克隆的vp1基因与已发表的其他vp1基因之间存在许多核甘酸差异.核甘酸的变异导致其编码蛋白质的某些氨基酸发生改变.氨基酸的改变主要集中在VP1蛋白的29、75、125、141、144、251、254、447位.在这些变异中,许多氨基酸的电荷和/或疏水性发生了改变,例如:Gly→Glu、Val→G1u、Ala→Thr、Leu→GIn、Cys→Trp、Leu→Arg、Arg→Ala、Gly→Ser、Ser→Ala、Glu→GIy、Gly→Thr等.通过CLUSTAL X软件比较了6个不同的vp1基因.该vp1基因已被(GenBank登录(登录编号:AF448446).VP1蛋白是CAV的唯一衣壳蛋白,因此VP1的氨基酸变异可能影响该蛋白质的抗原特征.对该vp1基因进一步进行免疫学研究具有重要意义,并且有可能应用该基因构建CAV基因工程疫苗.
A new vpl gene of chicken anemia virus ( CAV) was amplified via PCR and cloned into plasmid vector pUC18. The vpl gene contained 1 347 base pairs and the putative VP1 protein consisted of 449 amino acids. Comparisons of its sequence data with those of other vpl genes published in GenBank via DNA BLAST software showed that there were many nucleotide differences between the sequence of the cloned vpl gene and the others. The variations of nucleotides resulted in some changes of the amino acids encoded by this gene. The amino acid changes mainly concentrated on the loci, such as positions of 29, 75 , 125 , 141 , 144, 251, 254 , 447. Among them there were many amino acid changes which resulted in charge and/or hydropho-bicity changes, for examples, Gly→Glu, Val→Glu, Ala→Thr, Leu→Gln, Cys→Trp, Leu→Arg, Arg→Ala, Gly→Ser, Ser→Ala, Glu→Gly, Gly→Thr, and so on. Six different vpl genes were analyzed via CLUSTAL X software as well. The cloned vpl gene had been accessioned by GenBank ( Accession No. : AF448446) , and it is a new vpl gene. As VP1 protein is thought to be the only capsid protein of CAV, the amino acid changes may affect the antigenic character of VP1 protein encoded by this new vpl gene. It is very important for further immunological research on this new vpl gene, and it might be used to construct the genetic engineered vaccine against CAV. Fig 1, Tab 3 , Ref 20
出处
《应用与环境生物学报》
CAS
CSCD
2004年第2期189-193,共5页
Chinese Journal of Applied and Environmental Biology
基金
黑龙江省科学技术委员会项目(No.230102)~~
