摘要
目的对细粒棘球蚴中国大陆株14-3-3蛋白(E．g-14-3-3Pc)基因进行克隆及序列分析,为进一步重组抗原的表达奠定基础。方法根据互联网GeneBank中检索到的E．g-14-3-3Pc序列设计一对PCR 引物,用RT-PCR方法以细粒棘球蚴总RNA为模板扩增出目的DNA片段,并克隆于载体pGEM-T,测定其核苷酸序列。结果成功克隆出E．g-14-3-3Pc基因片段,测序证明克隆基因确为E．g-14-3-3Pc。结论成功构建细粒棘球蚴pGEM-E．g 14-3-3菌株。
Objective To construct a recombinant plasmid pGEM - E. g 14 - 3 - 3 for cloning and sequence analyzing, to establish a foundation for further study on the expression of E. g - 14 - 3 - 3 Proteins. Methods Specific primers were designed and synthesized according to published nucleotide sequence in the Genebank database. Coding region gene of E. g 14 - 3 - 3Pc was amplified by RT - PCR using Echinococcus granulosus RNA as template. The product from PCR was cloned into pGEM - T vector for screening and analyzing. Resttlts The region gene of E. g 14 - 3 - 3Pc was cloned successfully and was proved as certain E. g 14 - 3 - 3Pc by sequence analysis. Conclusion pGEM - E. g 14 - 3 - 3 was successfully constructed.
出处
《宁夏医学院学报》
2006年第2期93-95,98,共4页
Journal of Ningxia Medical College
基金
国家自然科学基金项目(30260105)
关键词
细粒棘球蚴
14-3-3蛋白
重组抗原
序列分析
echinococcus granulosus
14- 3- 3 protein
recombinant antigen
sequence analysis
