Vero细胞nucleolin和fibrillarin基因的克隆及序列分析

Cloning and sequence analysis of nucleolin and fibrillarin gene from Vero E6 cells

为克隆及分析Vero细胞的nucleolin和fibrillarin基因,本研究根据人类的相关基因设计两对引物,从Vero E6细胞中扩增nucleolin和fibrillarin基因。测序结果显示nucleolin基因序列长度为2 139 bp,编码712个氨基酸;fibrillarin基因序列长度为969 bp,编码322个氨基酸。与GenBank中登录的人类相应序列相似性高达97%,处于同一进化分支,表明该基因具有种属特异性并且保守性很高;同时利用软件对nucleolin和fibrillarin预测蛋白进行了生物信息学分析,从而为研究冠状病毒核蛋白与核仁蛋白相互作用奠定了基础。

To clone and analysis the genes of nucleolin and fibrillarin of Vero cells, the two genes was amplified by PCR with specific primers desigened according to human nucleolin and fibrillarin gene sequences in Gerd3ank. The results showed that the nucleolin contained a 2,136 bp ORF encoded 712 amino acids （aa）, the fibrillarin gene contained a 969 bp ORF encoded 322aa. Nucleotide sequence analysis showed that nucleolin and fibrillarin shared more than 97% homology and classified into the same evolutionary clade with that of human genes. The coloned genes would be facilitated for study on. Interaction of coronavirus N protein and nucleolus protein.