| 摘要: |
| 根据GenBank中报道的CCVInsavc-1株核蛋白基因(M)序列,用特异性引物对分离的CCVV1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连接得到重组质粒pTM,并进行了核苷酸序列测定。结果表明,该基因全长为789bp,编码263个氨基酸,其中前17个氨基酸为信号肽。与CCV标准毒株Insavc-1M基因相比,两者核苷酸的同源性为92.1%;推导的氨基酸序列同源性为90.9%,差异主要发生在该蛋白的N端前1/3区域内。在推导的M蛋白氨基酸序列中,发现了3个明显的疏水区,提示该蛋白可能是一个反复跨膜的蛋白;在N端15～17,38～40,234～236位氨基酸存在3个潜在的N-联糖基化位点,可能是形成该蛋白表面抗原决定基的位点。另外,推导蛋白氨基酸序列的疏水性和抗原表位与Insavc-1株M蛋白存在一定差异,提示两者免疫原性可能有差别。 |
| 关键词: 犬冠状病毒 膜蛋白基因 基因克隆 序列分析 |
| DOI： |
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| 基金项目:国家自然科学基金项目(30000123) |
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| Cloning and sequence analysis of membrane protein gene of canine coronavirus strain V1 |
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| Abstract: |
| According to membrane protein gene sequence of canine coronavirus strain V1 reported by GenBank,a pair of specific primers was designed and used to amplify M gene.The positive PCR product was purified and ligatured with pGEM-T.The correct positive recombinant was used for sequencing.The complete length of M gene of CCV V1 was 789 bp and encoded 263 amino acids.The initiative 17 amino acids was signal peptide.The homology of nucleic acid and amino acid between CCV V1 and Insavc-1 was 91.7% and 90.2% respectively.The different parts mainly exist in front of one third region of protein.Three hydrophobic regions exist in the protein,which denotes that it is a continuous cross-membrane protein.Three N-glycosylating sites were also found in region of 15-17,38-40 and 234-236 amino acids,which possibly consist of antigenic site on the surface of the protein.Moreover,there were some differences in the hydrophobicity and antigenic index between two strains,which may cause some difference in immunogenicity. |
| Key words: canine coronavirus membrane protein gene gene cloning sequence analysis |
