新城疫病毒F48E9株F基因主要功能区的核苷酸序列分析

Nucleotide Seqence Analysis of Main Functional Domain of NDV F48E9 Strain F Gene

本试验首先以ＮＤＶＦ４８Ｅ９株的基因组ＲＮＡ为模板，逆转录合成Ｆ基因ｃＤＮＡ第一链，再通过ＰＣＲ技术扩增Ｆ基因的ｃＤＮＡ，然后将其克隆到质粒ｐＵＣ１９中，经分子量比较、酶切分析、ＰＣＲ等方法证明，我们已经获得了ＮＤＶＦ４８Ｅ９株Ｆ基因的阳性克隆。经初步序列分析，ＦＡ段核苷酸序列与参考株（Ｄ２６／７６）序列同源性为８９％，ＦＢ段同源性为９１％。二者的氨基酸序列表明，Ｆ４８Ｅ９株Ｆ蛋白裂解位点与其它强毒株裂解位点氨基酸组成相同，即１１２ＲＲＱＲＲ１１６Ｆ１１７。这两段序列中包含的三个Ｃｙｓ残基和三个潜在的糖基化位点都相当保守。

cDNA ecoding the F gene of NDV F48E9 strain was cloned.At first,virion RNA was abstracted from purified NDV F48E9 strain and used as a template for cDNA synthesis.then cDNAs of F gene were amplified by PCR and ligated with the pUC19.The positive clones of F gene were identified by molecular weight,Restriction Endonucleases digestion and PCR.The result of preliminary sequence analysis indicated that FA gene prducts exhibited homology of 89% and FB homology of 91%.Cysteine residues and potential asparagine linked glycosylation sites are conserved.The same as other velogenic strains F48E9 strains consisted of two dibasic residues with a intervening glutamine, 112 R R Q R 116 R,and a phenylalaine(F) residue at position 117.