摘要
根据GenBank中收录的犬瘟热病毒(CDV)Onderstepoort株核蛋白基因(N)序列,设计了1对特异性引物,用该引物对分离的TN野毒株进行了RT PCR扩增;将扩增得到的PCR片段纯化后与pGEM T质粒连接,得到重组质粒pTN。经核苷酸序列测定,CDVTN株N基因的ORF全长1569bp,编码523个氨基酸;将TN野毒株与GenBank中收录的其他CDV毒株进行比较,N基因核苷酸序列的同源性为94%～99%,推导的N蛋白氨基酸序列的同源性为97%～99%。TN株与Onderstepoort疫苗株氨基酸序列的差异主要集中在N端(17～159位)和C端(408～519位),中间的区域则高度保守。此外,在CDVN蛋白N端281～289位上也发现存在与麻疹病毒(MV)N蛋白完全相同的Y P A L G L H E F9肽序列,推测可能是诱导CTL活性的靶蛋白的抗原表位之一。氨基酸组成分析发现,推导的N蛋白氨基酸序列中亮氨酸、丝氨酸和异亮氨酸所占比例很高,提示该蛋白具有高度螺旋和缠绕的分子结构。
According to the nucleocapsid (N) protein gene sequence of the canine distemper virus (CDV) strain Onderstepoort which was reported in GenBank database, a pair of specific primers was (designed) and used to amplify the Ngene of the CDV strain TN. The positive PCR product was purified and ligated with pGEM-T. The correct positive recombinant was used for sequencing. The open reading frame length of the Ngene of the CDV strain TN was 1 569 bp and encoded 523amino acids. The homology of nucleic acid and amino acid between the CDV strain TN and the other CDV strains published in GenBank ranged from 94% to 99% and from 97% to 99% respectively. The difference of nucleocapsidprotein (between) the TN strain and the Onderstepoort strain focused chiefly on both the 17-159 region and the 408-519 region. The region between the variable regions showed to be high conservative. Additionally, the (sequence) of Y-P-A-L-G-L-H-E-F owned by the N protein of measles virus (MV) was also found in the 281-289 region of the N protein of the CDV strain TN, which maybe acted the antigenic epitope recognized by CTL cells. The Leu , Ser and Ile amino acids have high content in the deduced N protein (amino) acid (sequence) from the N gene of the CDV strain TN, which indicated that the N protein exists complicated (molecular) structure such as helix and twine.
出处
《中国兽医科技》
CSCD
北大核心
2004年第4期24-28,共5页
Chinese Journal of Veterinary Science and Technology
