Cloning, prokaryotic expression and immunoreacticity of nodal modulator 1 gene of Taenia saginata asiatica
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Graphical Abstract
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Abstract
ObjectiveTo construct prokaryotic recombinantplasmids of nodalmodulator 1 gene of Taenia saginata asitica, to express and purify the recombinantprotein and to conduct prelininary inmunoreacticity study.MethodsApair of priners was designed according to the known sequence of NOMO1 gene The NOMO1 gene was amplified by PCR and the encoding sequence was cloned into the prokaryotic expression vector pET28a(+)and then expressed in E.coli BL21 with IPTGinduction.The recombinant protein was detected by SDS-PAGE Protein and the inmunoreacticity study was conducted with Western-blotting.ResultsThe recombinant plasmid pET28a(+)-Ta NOMO1 was successfully constructed SDS-PAGE results showed that the gene expression took place in Escherichia coliBL21/DE3, and highly pure protein was achieved after the dissolving, refolding and particle exchange chromatography of inclusion body deposits Western blotting anlysis of NOMO1 recombinant protein testified that the recombinant protein reacted with Taenia asiatica and Taeniarhynthus saginatus infected patient seaun,which indicated its inmunoreacticity.ConclusionThe NOMO1 gene is successfully cloned.The recombinant protein is obtained through expression and purification, and the gene's inmunoreacticity is confiuned,which provides basis for further studies of the gene.
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