新城疫病毒广西分离株M基因的克隆与序列分析

Cloning and Nucleotide Sequence Analysis of Matrix Protein Gene of Newcastle disease virus Isolates in Guangxi

根据基因库中新城疫病毒M基因核苷酸序列,设计一对特异性引物,对2000年-2003年期间广西分离的11个NDV毒株的M基因进行RT-PCR扩增、克隆和序列测定。结果表明,11个NDV广西分离株的M基因都为1223个核苷酸,含有1个1095bp核苷酸阅读框(ORF),编码364个氨基酸。序列分析结果表明,这11个NDV分离株核苷酸的同源性在86%～99.9%之间,推导氨基酸的同源性在89.3%～99.7%之间;11个毒株与国内外其他7个NDV毒株比较核苷酸的同源性在83.3%～98.5%之间,推导氨基酸的同源性在87.4%～98.6%之间。

A pair of specific primers were designed and synthesized according to Matrix（M） protein gene sequences of Newcastle disease virus （NDV） strains from GenBank. M genes of eleven NDV isolates from Guangxi outbreak ND chicken farms between 2000-2003 were amplified by reverse transcription-polymerase chain reaction （RT-PCR） technique. The full-length M genes of eleven NDV isolates in Guangxi were sequenced after the products of amplification being cloned. All complete nucleotide sequences of M gene of eleven NDV isolates in Guangxi are 1 223 bp long, including 1 095 hp ORF encoding 364 amino acids. The results of M gene analyses indicated that the homologies of the eleven NDV isolates＇ nucleotides are between 86%-99.9% and the homologies of the deduced amino acid sequences are between 89.3%-99.7%. Comparison of M protein complete sequences and the deduced amino acid sequences of eleven NDV isolates with seven other published NDV revealed that the homologies of the nucleotide are between 83.3%-98.5% and the homologies of the deduced amino acid sequences are between 87.4 %-98.6 %.