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目的构建弓形虫微线体蛋白MIC3的真核表达质粒,为进一步研究MIC3蛋白功能奠定基础。方法 PCR扩增弓形虫MIC3目的基因,将纯化的目的基因插入到真核表达质粒PCDNA3.0,并转入DH5α宿主菌中,在含有青霉素的SOB平板上筛选阳性重组子,采用酶切和PCR扩增方法对重组质粒进行鉴定。结果重组质粒PCDNA-MIC3经单双酶切及PCR扩增都得到以原插入片段MIC3基因1120bp大小相同的片段。结论成功构建弓形虫MIC3基因真核表达质粒PCDNA-MIC3.
Objective To construct the eukaryotic expression plasmid of microtubule protein MIC3 of Toxoplasma gondii and lay the foundation for further study on the function of MIC3 protein. Methods The target gene of MIC3 of Toxoplasma gondii was amplified by PCR. The target gene was inserted into the eukaryotic expression plasmid PCDNA3.0 and transformed into DH5α host. The recombinant plasmid was screened on the SOB containing penicillin. Amplification method to identify recombinant plasmids. Results The recombinant plasmid PCDNA-MIC3 was obtained by single-enzyme digestion and PCR amplification. The fragment of 1120bp in size of the MIC3 gene of the original insert was obtained. Conclusion The eukaryotic expression plasmid pcDNA-MIC3 of MIC3 gene of Toxoplasma gondii was successfully constructed.