猪嵴病毒CH441株VP1基因的克隆与序列分析

Molecular Cloning and Sequence Analysis of the VP1 Gene of Porcine kobuvirus

为了深入研究嵴病毒（swKoV）主要结构蛋白基因 VP1，根据 GenBank中已发表的猪嵴病基因序列设计特异性引物，采用 RT-PCR方法扩增猪嵴病毒 CH441株 VP1基因，并对其进行克隆与测序分析。结果表明，swKoV CH441株的 VP1基因为762 bp，与 GenBank已发表的嵴病毒属的15株嵴病毒序列的 VP1基因相比较，swKoV CH441株的VP1基因与其他各毒株 VP1基因的核苷酸同源性为81.5%~90.2%，氨基酸同源性为86.6%~96.9%，进化分析显示，swKoV CH441株与 GS-1株之间的亲缘关系较近。生物信息学分析显示，VP1蛋白理论等电点（ pI）为4.40，理论分子质量为26.9782 kDa；其序列上共发现18个磷酸化位点，分别为Ser（7）、Thr（6）和Tyr（5），而蛋白的磷酸化与信号转导有关，预测该蛋白为一重要的信号转导分子；无信号肽和跨膜区。为进一步开展 swKoV CH441株 VP1基因在遗传变异等方面的研究奠定了理论基础。

The aim of the study to investigate the main structural protein of the Kobuvirus VP1 gene. According to the sequences of PKV deposited in GenBank,a pair of special primers was designed for amplifying the VP1 gene of swKoV CH441 strain by RT-PCR. The results of sequence analysis showed that the whole VP1 gene of swKoV CH441 strain consisted of 762 bp. Compared with 15 PKV strains which were deposited in GenBank,the homology of nucleotide sequences was 81. 5%~90. 2%,and the homology of deduced amino acids was 86. 6%~96. 9%. Evo-lution analysis indicated that the swKoV CH441 strain was closely related to GS-1 strains. The bioinformatics analy-sis demonstrated that the isoelectric point and molecular weight of non-structural protein VP1 were 4. 40 and 26. 978 2 kDa. The protein had no signal peptide and transmembrane domain. There were 18 phosphorylation sites including 7 Sers,6 Thrs and 5 Tyrs. Protein phosphorylation was concerned with signal transduction,so this protein may be a signaling molecule. The results provided a theoretical foundation for further research on the study of VP1 gene（ protein）in the genetic variation.