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新城疫病毒V4株NP和P基因及其间隔区序列的克隆与分析
刘芳宁1,2, 闫丽辉2, 曹殿军2
1.西北农林科技大学 动物科技学院;2.中国农业科学院 哈尔滨兽医研究所
摘要:
采用RTPCR技术克隆了新城疫病毒(NDV)V4株长约4200bp的核背酸序列,并进行了序列测定与分析。结果表明,该序列中包含有NDVV4株NP基因编码区、NP和P基因间隔区以及P基因的全部核苷酸序列。NDVV4株NP和P基因与其他8个NDV毒株相应部分进行同源性比较。与Quuesland V4株的同源性最高。NDVV4株NP和P基因间隔区序列与其他13株NDV毒株相应部分的比较结果表明,F48E9等强毒株较Lasota等弱毒、无毒毒株和试验用V4株在这个区域多了6个碱基,这6个碱基插入的位置在Lasota等弱毒、无毒毒株全基因组序列的1 647~1 648 nt处;参比的14株NDV在这个区域的同源性为63.9%~100%,弱毒株之间的同源性较高,弱毒株与强毒株之间及强毒株之间的同源性差异较大。
关键词:  新城疫病毒  NP基因  P基因  NP和P基因间隔区序列  克隆  序列分析
DOI:
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基金项目:国家重点基础研究发展规划(973)项目(G1999011902);黑龙江省科学技术计划项目(GB02B505)
Cloning and sequencing of NP gene,P gene and intergenic region in NP and P gene coding region junction of Newcastle disease virus V4 strain
Abstract:
Utilizing RT-PCR,the cDNA fragment of NDV V4 strain was successfully amplified,which was about 4.2 kb in length,and was cloned and sequenced.The cDNA fragment covers the open reading frame (ORF),the intergenic regions in NP and P gene coding region junction and the whole nucleotide sequence of Phosphoprotein (P) gene.According to the comparative analysis between the nucleotide and amino sequence of NP and P gene in NDV V4 and the counterpart in 8 other NDV strains,the homology was highest with Queenland V4.Comparison of nucleotide sequence was carried out between intergenic region in NP and P gene coding region junction of NDV V4 strain and the counterpart of other 13 NDV strains respectively.The results showed that the mesogenic or velogenic strains,such as F48E9 and ZJ1,had the extra 6 nucleosides insertion between 1 647 nt and 1 648 nt of lentogenic strains,such as La Sota and D26,indicating the homology was different (63.9%-100%),and presented the homology between lentogenic strains was higher than that between velogenic strains,and than that between lentogenic strains and velogenic strains.
Key words:  newcastle disease virus (NDV),nucleocapsid protein(NP),phosphoprotein (P),intergenic regions in NP and P gene coding region junction,cloning,sequencing