弓形虫MIC3基因真核表达质粒的构建与鉴定

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DH5ct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．

Construction and identification of Eukaryotic Expression Plasmid of Gene MIC3 of Toxoplasma Gondii

Objective To construct eukaryotic expression plasmid of microneme protein MIC3 of toxoplasma gon- dii, so as to lay a foundation for further study the function of protein MIC3. Methods The target gene of toxo- plasma gondii amplified by PCR, and the purified target gene was inserted into eukaryotic expression plasmid PCDNA3.0 and then transferred into host bacteria DH5ot. The positive recombinant was screened from SOB plate that contains penicillin, and then the recombinant plasmid was identified by enzyme digestion and meth- od of PCR amplification. Results For recombinant plasmid PCDNA - MIC3 was amplified to obtain fragment that equaled the size of the originally inserted fragment of gene MIC3 1120bp after single or double enzyme di- gestion or PCR cation. Conclusions Eukaryotic Expression Plasmid of Gene MIC3 of Toxoplasma Gondii was successfully constructed.