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.
JIANG Tao, ZHANG Donglin, NIE Hao, ZHAO Junlong, YAO Baoan
. Construction and expression of the eukaryotic
expressed plasmid of gene
from in IBRS-2
cells[J]. Frontiers of Agriculture in China, 2008
, 2(4)
: 498
-501
.
DOI: 10.1007/s11703-008-0075-6
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