南非2型口蹄疫病毒VP1基因的核苷酸及其氨基酸序列分析

Sequence analysis of the nucleotide and amino acid of VP1 gene of South African Territories type 2 Foot- and- mouth disease virus

VP1基因的遗传衍化关系是口蹄疫病毒分型的依据,而且许多重要的抗原位点也都位于VP1蛋白上。为了对南非2型口蹄疫病毒(SAT2-FMDV)VP1的编码基因序列及其氨基酸序列进行分析,为南非2型口蹄疫的诊断及疫苗设计提供理论基础,试验从NCBI下载大量的南非1/2/3型口蹄疫病毒(SAT1/2/3-FMDV)的1D2A2B基因片段,通过序列分析选择相似性较低的序列代表SAT1/2/3,将选中的代表序列连接于p UC57载体上进行合成,再对合成的含目的序列的穿刺菌进行划板、挑单克隆、摇菌、提质粒,得到重组质粒初步进行酶切鉴定后进行测序,测序正确后采用DNAStar软件包对这些合成序列进行分析。结果表明:SAT2-FMDV VP1的编码基因核苷酸序列变异度较大;虽然SAT2口蹄疫病毒VP1蛋白上主要抗原位点位置与其他血清型的差异性并不大,但是关键氨基酸已发生变化。

Genetic derivation relationship of VP1 gene is the basis for typing Foot - and - mouth disease virus （ FMDV）, and most of important antigen sites were located in the VP1 protein. To analyze the coding gene sequence and amino acid sequence of VP1 gene of South Africa Territories type 2 foot and mouth disease virus（ SAT2 - FMDV）, and provide a theoretical basis for the diagnosis and vaccine design of SAT2 - FMDV, a lot of 1D2A2B gene fragments of SAT1/2/3 - FMDV were downloaded from NCBI. The sequences with low similarity were selected to represent the SAT1/2/3 by sequence analysis,respectively. The selected representative sequences were connected to pUC57 vector for synthesis, and then puncture bacteria which contained target sequences were coated to plates, followed by selecting monoclonal colony, shaking bacteria and extracting plasmid. The constructed recombinant plasmid was preliminarily identified by enzyme digestion, and then was used for sequencing. Once confirmed, these synthesized sequences were analyzed by DNAstar software. The results showed that there is greater variabili- ty in the nucleotide sequence of the coding gene of VP1 of SAT2 - FMDV. Although there was no significant difference in the location of main antigen sites between VP1 protein of SAT2 - FMDV and the other serum types, the key amino acids has been changed.