一株猴泡沫病毒野毒Pol基因的cDNA的直接测序

The cDNA Nested-PCR Direct Sequencing pol Region of A Simian Foamy Virus in Rhesus Monkey

本研究对一株来源于医学灵长类中心 (CentreforMedicalPrimates)大笼饲养的恒河猴的原代猴肾细胞 (RhesusMonkeykidney ,RMK) ,377细胞株 ,经细胞培养三周后 ,具有典型泡沫样病变 ,即CPE( + )的细胞 ,进行Pol基因 ( 5898- 6362 )的cDNA的Nested -PCR扩增 ,对产物位置 5898- 6343区域进行直接测序。结果证明 :该株细胞所带的SFV与GENEBANK中的SFV - 1在该区域有 48个核苷酸存在差异 ( 1 0 .78% ) ,与SFVmac在该区域有 45个核苷酸存在差异 ( 1 0 .1 1 % )。VspⅠ酶切实验结果与之吻合 ,显示出该前病毒株的PCR产物与SFV - 1在该片段的第 69核苷酸有变异 ,碱基由T→A。

A 377 Kunming cell line of Rhesus Monkey Kidney (RMK) from Centre for Medical Primates was amplified the cDNA fragments of the pol gene by Nested-PCR,which of a natured infected SFV showed typical CPE after cultured for three weeks,sequences of products position(nt 5989-6343)were determined directly.The result indicated that the nucleotid sequences in the region had 48 nucleotids difference(10.78%) between SFV infected cell and SFV-1 in GENE BANK,and there was 45 nucleotides difference(10.11%) compared with SFVmac in the region, the test with VspⅠ showed aligment. The paper have a significant that the nucleiotides sequences was determined directly with the Nested-PCR products in molecular epidenicology,it will supply a rapid、sensitive and reliable cDNA probe for diagnosing agent Kit of nucleotids.