新城疫病毒F48E9株及东北地区流行株F基因遗传变异分析

采用异硫氰酸胍／酚／氯仿抽提一步法，提取新城疫病毒（ＮＤＶ）Ｆ４８Ｅ９株及２个东北地区分离株ＤＢ３、ＤＢ５基因组ＲＮＡ，并以其为模板经反转录合成Ｆ基因ｃＤＮＡ第１链，再利用ＰＣＲ技术分段增出Ｆ基因的ｃＤＮＡ。Ｆ４８Ｅ９、ＤＢ３和ＤＢ５株Ｆ基因的ｃＤＮＡ经酶切修饰后，插入ｐＵＣ１９的多克隆位点中，转化感受态Ｅ。ｃｄｉ ＤＨ５ａ，经相对分子质量比较、酶切分析及ＰＣＲ等鉴定方法筛选出Ｆ４８Ｅ９、ＤＢ３和

Genetic Variance Analysis of F Gene of NDV F48E9 Strain and Isolates from Northeast China

Virion RNA were abstracted from purified NDV F48E 9, DB3 and DB5 strain, and used as templates for cDNA synthesis, the cDNA of F g ene were amplified by PCR and ligated with the pUC19. The positive clones of F g ene of NDV F48E9, DB3 and DB5 strain were identified by molecular weight, restri ction endonucleases digestion and PCR. The positive clones were sequenced by San g er′s method. Comparison and analysis of the nucleotide sequence and deduced ami no acid sequence with that of other 11 strains previously published was performe d by computer with DNASIS and MEGA software. The results showed that a high degr ee of functional and structural constraint exerted on the F glycoprotein. Howeve r, there were amino acid residues variations in coding region of f gene. Phyloge netic tree of the nucleotide sequence of NDV strains F gene was constructed by c omputer. The F48E9 and DB3 strains were found to be able to construct a new grou p independent from classic A,B,C groups, and the DB5 strain should belong to gro up B.