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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front. Agric. China    2008, Vol. 2 Issue (4) : 498-501     DOI: 10.1007/s11703-008-0075-6
Construction and expression of the eukaryotic expressed plasmid of gene from in IBRS-2 cells
JIANG Tao1, ZHANG Donglin2, NIE Hao2, ZHAO Junlong2, YAO Baoan3
1.State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University;College of Animal Science, Yangtze University;College of Veterinary Medicine, Huazhong Agricultural University; 2.State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University;College of Veterinary Medicine, Huazhong Agricultural University; 3.College of Veterinary Medicine, Huazhong Agricultural University;
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Abstract The sequence encoding MIC3 was obtained by amplification from genomic DNA of Toxoplasma gondii RH strain and cloned into the vector pMD18-T. The target 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 containing pcMIC3 plasmids were obtained under the selection of G418. The expressed proteins from the positive cells were detected by SDS-PAGE, Western blot and ELISA. The results showed that the DNA sequence identity 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.
Issue Date: 05 December 2008
 Cite this article:   
ZHANG Donglin,YAO Baoan,NIE Hao, et al. Construction and expression of the eukaryotic expressed plasmid of gene from in IBRS-2 cells[J]. Front. Agric. China, 2008, 2(4): 498-501.
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