弓形虫GJS株表面抗原1基因的克隆及真核表达质粒的构建  

Cloning and eukaryotic expression plasmid construction of the surface antigen 1 gene of Toxoplasma gondii GJS strain

作　　者：曹丽艳[1,2] 张德林[2] 张燕丽[1,2] 芦赟[1,2] 王艳华[2] 苟惠天[1,2] 付宝权[2] 赵晋军[1] 

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070 [2]中国农业科学院兰州兽医研究所,家畜疫病病原生物学国家重点实验室,甘肃省动物寄生虫病重点实验室,甘肃兰州730046

出　　处：《甘肃农业大学学报》2009年第3期34-38,共5页Journal of Gansu Agricultural University

基　　金：国家科技支撑计划项目(2007BAD40B05);公益性行业(农业)科研专项经费项目资助(200803017);甘肃省重大专项科技项目(2GS063-A43-013)

摘　　要：根据GenBank数据库中的RH株弓形虫表面抗原1(SAG1)基因序列设计1对引物,应用PCR技术从弓形虫GJS株速殖子基因组DNA中扩增编码SAG1的基因片段.PCR产物经纯化、酶切后定向插入pcDNA3.1(+)真核表达载体,转化至大肠杆菌DH5α,经酶切及PCR鉴定后测序,并进行序列分析.结果表明:从弓形虫GJS株基因组DNA中扩增出1 008 bp的特异SAG1片段,大小与预测值相符,与已知的SAG1基因核苷酸序列(GenBank登录号:S76248)的ORF和氨基酸序列的同源性均为99%.One pair of specific primers based on the published surface antigen 1 gene of Toxoplasma gondii RH strain sequence in GenBank was designed. The gene coding for surface antigen 1 （SAG1） was amplified from the genomie DNA of Toxoplasma gondii GJS strain by PCR. The amplified product was pu- rified, digested and inserted into pcDNA3. 1 （＋） eukaryotic expression vector digested with restriction enzymes. Then the recombinants were transferred into E. coli DH5a and identified by PCR, restriction enzyme digestion and sequencing. Sequence analysis indicated that the specific fragments of SAG1 gene isolated from genomic DNA of To3coplasma gondii GJS strain was 1 008 bp. Homology analysis showed that the ORF of nucleotide sequences and amino acid sequences of SAG1 shared 99% identity with those of the published Toxoplasma gondii SAG1 sequence （S76248）.

关 键 词：弓形虫 表面抗原1基因 克隆 真核表达质粒 

分 类 号：P382.5[天文地球—地球物理学]