Cloning and expression of MxA gene from chickens in E.coli and identification of biological activity of MxA
- VernacularTitle:鸡MxA基因的克隆、表达及生物学活性检测
- Author:
Lei CHEN
;
Guotuo JIANG
;
Weishan CHANG
- Publication Type:Journal Article
- Keywords:
Chickens MxA gene;
Gene clone;
Protein expression;
Biological activity
- From:
Chinese Journal of Immunology
2000;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To clone chickens MxA gene, construct its recombinant expression plasmid and induce the expression of fusion protein using a prokaryotic expression system.Methods:The MxA gene fragment was amplified by RT-PCR from CEF cells and subcloned into the pMD18-T vector, filtrated the positive clone and reclaimed the MxA.Subcloned the MxA into the prokaryotic expression plasmid pGEX-6p-1. After recombinant plasmid was induced by IPTG, the expressed proteins were analyzed by SDS-PAGE, the NDV intervence experiment and VSV-CEF restrain experiment.Results:The sequence of MxA gene amplified by RT-PCR was the same as the sequence in gene map of Genebank; SDS-PAGE, the NDV intervence experiment and VSV-CEF restrain experiment showed that a protein was expressed, the molecular weight of this protein was 45 000, which was the same as the fusion protein GST-MxA.Conclusion:The MxA is cloned and its recombinant expression plasmid is constructed successfully.The fusion protein GST-MxA is successfully expressed in the prokaryotic expression system E.coli DH5? induced by IPTG. This research lay a foundation for further studying on ints antiviral effects and exploring new way of antiviral medication.
