| 摘要: |
| 【目的】对鸽源新城疫病毒（NDV）陕西分离株P基因进行克隆和序列分析，为研究P基因及V基因的功能奠定基础，并为防治新城疫提供科学依据。【方法】参考GenBank发表的NDV基因序列设计1对特异性引物，RT-PCR扩增NDV/Pigeon/ShX/China/2003 P基因的全长片段，并进行克隆、序列测定和分析。【结果】NDV/Pigeon/ShX/China/2003 P基因全长1 451 bp，其完整的ORF长度为1 188 bp，编码395个氨基酸。同源性比较可知，NDV/Pigeon/ShX/China/2003与其他分离株P基因的核苷酸序列同源性在63.6%～92.9%，推导的氨基酸序列同源性在67.7%～92.2%。系统发育进化树表明，NDV/Pigeon/ShX/China/2003与鸽源分离株P68亲缘关系最近，与F48E9和La Sota毒株处于进化树的不同分支。【结论】NDV/Pigeon/ShX/China/2003与国内外已报道的NDV毒株间存在较大变异。 |
| 关键词: 新城疫病毒 P基因 序列分析 |
| DOI： |
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| 基金项目:国家“十一五”科技支撑计划子课题(2006BAD06A08) |
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| Cloning and sequence analysis of P of NDV strains isolated from pigeon of Shaanxi |
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| Abstract: |
| 【Objective】 In order to provide scientific basis for the prevention and control of outbreak and epidemic of Newcastle disease (ND),the Pgene of Newcastle disease virus (NDV) isolated from Pigeon of Shaanxi (NDV/Pigeon/ShX/China/2003) was amplified and the sequence was analyzed.【Method】 The virus was proliferated by inoculating into the non-immune chicken embryos,and was identified by HA and HI method.A pair of specific primers were designed according to the gene sequence of NDV published in GenBank,and was used to amplify the fragment of P gene (1 451 bp) by RT-PCR method,which was cloned and sequenced.【Result】 The P gene of NDV/Pigeon/ShX/China/2003 contains 1 451 bp and the open read frame (ORF) contains 1 188 bp,which encodes a polypeptide of 395 amino acids.Nucleotide sequences and deduced amino acids sequences analysis showed:homologous rate of P gene nucleotides and deduced amino acids sequences was 63.6%-92.9% and 67.7%-92.2% respectively.The phylogenetic tree showed that there was a close genetic relationship between the Shaanxi isolate and P68 strain.However NDV/Pigeon/ShX/China/2003,F48E9 and La Sota strains were on the different branches of the phylogenetic tree.【Conclusion】 The result indicated that there was evident variation between NDV/Pigeon/ShX/China/2003 and between strains isolated from domestic and abroad. |
| Key words: Newcastle disease virus P gene sequence analysis |
