猪瘟病毒中国标准强毒株——F114株全长cDNA的构建  

CONSTRUCTION OF THE FULL-LENGTH cDNA CLONE OF CHINESE VIRULENT STRAIN—F114

作　　者：聂玉春[1] 柯叶艳[1] 陈建国[1] 丁明孝[1] 

机构地区：[1]北京大学生命科学学院,北京100871

出　　处：《微生物学报》2001年第4期452-456,共5页Acta Microbiologica Sinica

基　　金：国家攀登计划资助项目! (No .85 4 4 0 2 0 5);国家重点基础研究发展规划项目! (G1 9990 1 1 90 4 )资助&&

摘　　要：应用RT PCR技术 ,分段克隆了猪瘟病毒中国标准强毒株F1 1 4株的全基因组 ,经测序验证 ,用DNAman软件比较分析与其它几个代表毒株的同源性 ,发现与Brescia、Alfort及C株核苷酸序列同源性分别为 96 80 % ,86 0 3%和 95 70 % ,氨基酸序列同源性分别为 98 54% ,93 33%和 97 41 %。将各段cDNA连接到pGEM T和pBluescriptⅡsk+质粒 ,得到两个亚克隆sk 0 1 64(即sk+质粒中插入片断为 1～ 6441nts,依次类推 )和sk 641 2 2 ,再将两个亚克隆连接成sk 1 2 2 97,即CSFV全长cDNA克隆。Seven subclones covered the complete genome of classical swine fever virus Chinese virulent strain F114 were obtained by reverse transcription PCR. The complete nucleotide sequence of the genome of strain F114 was determined by sequencing.The cDNA fragments were then assembled and inserted downstream of a T7 promoter in pBluescript Ⅱ sk + plasmid vector to obtain the full\|length cDNA clone sk\|12297.Homology comparison of the nucleotide and amino acid sequence of strain F114 with the known sequences of HCLV,Brescia and Alfort showed 95.70%,96.80%,86.03% identity in nucleotide and 97.41%,98.54%,93.33% identity in amino acid respectively.

关 键 词：猪瘟病毒 全长CDNA 序列同源性 

分 类 号：S855[农业科学—临床兽医学]