新城疫病毒F_(48)E_8株HN基因的克隆和序列分析及与国外NDV HN基因的同源性比较

Cloning and Sequencing of HN Gene of Newcastle Disease Virus (NDV) Strain F_(48)E_8 and Homology of It to NDV HN Genes Abroad

目的 克隆ＮＤＶ Ｆ４８Ｅ８株ＨＮ基因，与国外ＮＤＶ ＨＮ基因进行比较分析。方法 提取 ＲＮＡ，应用 ＲＴ－ＰＣＲ一次性扩增出ＮＤＶ Ｆ４８Ｅ８株的全长ＨＮ基因，将该ＨＮ基因插入ｐＫＳ（－）后进行克隆并测序。结果 ＮＤＶ Ｆ４８Ｅ８株的ＨＮ基因核苷酸长度为１７１３ｂｐ，编码５７１个氨基酸，有５个糖基化位点，１２个极保守的半胱氨酸残基，与 国外发表的ＮＤＶ序列相符。结论ＮＤＶＦ１８Ｅ８株ＨＮ蛋白基因与国外发表的ＮＤＶ ＨＮ蛋白基因核苷酸同源性在 ８３．３％～８９．２％之间，推导的氨基酸同源性在８７．６％－９１．１％之间。

Objective To clone FIN gene of Newcastle disease virus(NDV) strain F48E8 and compare it with NDV TIN genes abroad. Methods After the RNA of NDV strain F48E8was extracted, a whole length of HN gene was amplified by RT-PCR, then inserted into vector pKS(-)for cloning, and identified by sequencing. Results The whole length of the cloned gene was 1713bp. It contained 5 glycosylation sites and 12 highly conservative cysteine residues,and could encode 571 amino acids. All the results were consistent with those of NDV HN gene abroad. Conclusion Compared with the HN gene sequence abroad,the homologies of the clone HN gene and deduced amino acid sequence were 83.3% to 89.2% and 87.6% to 91.1% respectively.