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目的:构建弓形虫致密颗粒抗原GRA7的真核重组表达质粒。方法:设计GRA7的特异引物,采用多聚酶链反应(PCR)技术从弓形虫RH株基因组DNA中扩增编码GRA7的基因片段,经克隆至pMD18-T载体后,亚克隆至真核表达载体pVAX1而构建真核重组表达质粒pVAX1-GRA7。结果:PCR扩增出GRA7基因的特异片段,所获克隆的序列正确,并被亚克隆到真核表达载体pVAX1,构建了真核重组表达质粒pVAX1-GRA8。结论:成功构建了GRA7的真核重组表达质粒pVAX1-GRA7。
Objective: To construct eukaryotic recombinant plasmid of Toxoplasma gondii GRA7. Methods: The specific primers of GRA7 were designed. The gene encoding GRA7 was amplified from genomic DNA of RH strain Toxoplasma gondii by polymerase chain reaction (PCR). The fragment was cloned into pMD18-T vector and subcloned into eukaryotic expression vector pVAX1 Eukaryotic recombinant plasmid pVAX1-GRA7 was constructed. Results: The specific fragment of GRA7 gene was amplified by PCR. The cloned sequence was correct and subcloned into eukaryotic expression vector pVAX1 to construct eukaryotic recombinant plasmid pVAX1-GRA8. Conclusion: The eukaryotic recombinant plasmid pVAX1-GRA7 of GRA7 was successfully constructed.