细粒棘球蚴中国大陆株重组14-3-3蛋白基因的克隆和序列分析

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.g14-3-3菌株.

Cloning and Sequence Analysis of A Recombinant 14-3-3 Gene on Echinococcus Granulosus of Chinese Mainland Strain

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.