作 者：
乔贞贞;秦艳红;乔奇;张德胜;田雨婷;高洁;张振臣
单 位：
吉林农业大学农学院;河南省农业科学院植物保护研究所
关键词：
甘薯卷叶病毒;基因组序列;外壳蛋白基因;原核表达
摘 要：
为了制备甘薯卷叶病毒(SPLCV)的特异性抗体,克隆了甘薯卷叶病毒江苏分离物SPLCV(JS∶XZ∶3-2)的全长基因,并对其基因结构及分子变异情况进行分析,同时对外壳蛋白(CP)基因进行了克隆和表达。利用PCR方法扩增并克隆了SPLCV(JS∶XZ∶3-2)的全基因组,测序分析表明,SPLCV(JS∶XZ∶3-2)基因组全长为2 834bp,含有6个开放阅读框架,与已发表的SPLCV其他分离物相比,全基因组核苷酸序列相似性为82.7%～94.4%。其中,CP基因由765个核苷酸组成,共编码254个氨基酸残基。SPLCV CP基因核苷酸序列与其他分离物的相似性在89.2%～90.6%,其中与SPLCV美国分离物(HQ333135)的相似性最高,为90.6%;与日本分离物(AB433788)的相似性最低,为89.2%;与中国大陆分离物(FJ515896)CP基因的核苷酸序列相似性为90.2%。将SPLCV CP基因克隆到原核表达载体pGEX-4T-3上,获得重组质粒,经IPTG诱导表达、SDS-PAGE分析得到了大小约为54.3kD的融合蛋白条带,说明CP基因在大肠杆菌中得到了高效表达。
译 名：
Sequencing of Sweet Potato Leaf Curl Virus Genome and Expression of Coat Protein Gene in E.coli
作 者：
QIAO Zhen-zhen1,QIN Yan-hong2,3,QIAO Qi2,3,ZHANG De-sheng2,3, TIAN Yu-ting2,3,GAO Jie1,ZHANG Zhen-chen2,3*(1.Faculty of Agronomy,Jilin Agricultural University,Changchun 130000,China; 2.Institute of Plant Protection,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China; 3.Henan Key Laboratory of Crop Pest Control,Zhengzhou 450002,China)
关键词：
sweet potato leaf curl virus(SPLCV);complete genomic sequence;coat protein gene;prokaryotic expression
摘 要：
The complete genome of sweet potato leaf curl virus(SPLCV) isolate JS∶XZ∶3-2 from Jiangsu province was amplified by PCR.Analysis of the genomic sequence showed that the genome of SPLCV(JS∶XZ∶3-2)was composed of 2 834 nt and included six open reading frames(ORFs).Compared with other isolates previously reported,the nucleotide sequence identity of the complete genome was 82.7%-94.4%.The CP gene consisted of 765 nt and encoded 254 amino acid residues.The nucleotide sequence of CP gene was 89.2%-90.6% identical to other isolates previously reported.The highest identity was 90.6% with the USA isolate(HQ333135) and the lowest identity was 89.2% with the Japan isolate(AB433788).The nucleotide sequence of CP gene was 90.2% identical to the China isolate(FJ515896).The CP gene was cloned into the expression vector pGEX-4T-3 for over-expression in prokaryotic cells.The SDS-PAGE result showed that a specific fusion protein about 54.3 kD was produced after induction by IPTG.
