,Construction and expression of the eukaryotic expressed plasmid of MIC3 gene from Toxoplasma gondii

• 作 者: Tao JIANG Donglin ZHANG Hao NIE Baoan YAO Junlong ZHAO
• 机 构: State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan 430070, C
• 出 处: 中国高等学校学术文摘·农学
• 发表日期: 2008年4期
• 关键词: Toxoplasma gondii MIC3 gene clone eukar-yotic expression

The sequence encoding MIC3 was obtained by amplification from genomic DNA of Toxoplasma gondii RH strain and cloned into the vector pMD18-T. The tar-get gene was subcloned into the eukaryotic vector pcDNA3.1 after the identification of pMD18-T-MIC3 by enzyme digesting, PCR amplification and sequencing. Then the target recombinant plasmids pcMIC3 were transfected into IBRS-2 cells, and the positive cells con-taining pcMIC3 plasmids were obtained under the selec-tion of G418. The expressed proteins from the positive cells were detected by SDS-PAGE, Weste blot and ELISA. The results showed that the DNA sequence iden-tity was 99.9% between amplified MIC3 and that from GenBank. The molecular weight of the recombinant MIC3 protein with good immuno-activity was about 39.2 ku. These available data would lay the foundation for further studies on DNA vaccine against Toxoplasma gondii.