Construction of Eukaryotic Expression Vector for Myospalax fontanierii Zona Pellucida 3 Gene (mZP3) and Its Expression in Chinese Hamster Ovary (CHO) Cell
Abstract：Zona pellucida 3 (ZP3) is the most important acceptor in the sperm-egg binding. In order to construct Myospalax fontanierii mZP3 eukaryotic expression vector pEGFP-mZP3 and detect the expression, mZP3 fragment was obtained by PCR and inserted into the eukaryotic expression vector pEGFP-N1. pEGFP-mZP3 was transfected to Chinese hamster ovary (CHO) cell and injected into Kunming mice (Mus musculus), its expression was detected using RT-PCR and Weston bolt. Identification result showed that expression vector pEGFP-mZP3 was constructed successfully, the transfected cells emitted green fluorescence. RT-PCR and Western bolt test results showed that mZP3 expressed in CHO cells, and the target genes were also detected in the liver of mice. These results suggested that expression vector pEGFP-mZP3 successfully expressed in CHO cells, and provide the basis for the control of Myospalax fontanierii by immunization approach.
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