日本血吸虫(大陆株)Sj26基因的克隆与序列分析  

Cloning and Sequence Analysis of Sj26 Gene of Schistosoma Japonicum(Chinese Strain)

作　　者：罗金萍[1] 陈喜圭[1] 沈定文[1] 李雍龙[2] 

机构地区：[1]咸宁学院医学院寄生虫学教研室,湖北咸宁437100 [2]华中科技大学同济医学院病原生物学系

出　　处：《咸宁学院学报（医学版）》2004年第5期323-325,共3页Journal of Xianning Univarsity(medical Sciences)

摘　　要：目的构建日本血吸虫 (大陆株 ) 2 6kDa谷胱甘肽 S 转移酶 (Sj2 6 )基因的真核表达载体 ,并测定基因序列和进行序列分析。方法用PCR的方法特异性地扩增Sj2 6基因 ,并将此基因克隆于真核表达载体pcDN3,转化大肠杆菌DH5α感受态细胞 ,阳性克隆的重组质粒DNA经酶切、PCR扩增鉴定 ,用双脱氧链末端终止法进行序列测定 ,应用BLAST方法分析Sj2 6基因序列并进行同源性比较。。结果PCR扩增得到特异的Sj2 6基因 ,双酶切及PCR鉴定表明获得正确的pcDNA3 Sj2 6重组质粒 ,测序结果获得的基因片段大小为 6 5 1bp ,编码 2 17个氨基酸残基。基因序列同源性为 99%。结论成功构建真核表达重组质粒pcDNA3 Sj2 6。Objective To construct a eukaryotic expression plasmid containing a gene encoding 26kDa glutathione s-transferases(Sj26),determine the sequence of Sj26 gene and compare with the sequence of Sj26 genes.Methods Sj26 gene was amplified by PCR.By cloning target gene into a eukaryotic expression pcDNA3,a recombinant plasmid pcDNA3-Sj26 was constructed and transferred into E.coli DH5α.The positive recombinant pcDNA3-Sj26 was screened and idengified by endonuclease digestion and PCR technique.The correct recombinant plasmid used as template,and the nucleotide sequence of Sj26 gene was determined by the dideoxy chain termination method.Using BLAST to analyze the sequence of Sj26 gene and the gene homology.Results Sj26 gene was specifically amplified.The correct recombinant plasmid pcDNA-Sj26 was constructed.The result of sequencing the nucleotide acids showed that the Sj26 gene was 651 base pairs,encoding 217 amino acid residues.The homology of nucleotide acid sequence encoding Sj26 gene was 99%.Conclusion The pcDNA3-Sj26 recombinant plasmid was successfully constructed.

关 键 词：日本血吸虫 基因序列 真核表达载体 PCDNA3 重组质粒 DH5Α PCR 阳性克隆 酶切 感受态细胞 

分 类 号：R383.24[医药卫生—医学寄生虫学] Q78[医药卫生—基础医学]