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- W2356390496 abstract "Objective To clone the blaNDM-1 gene of Acinetobacter baumannii,analyze the bioinformatics of NDM-1 gene and protein,then express and purify recombinant NDM-1 protein.Methods The blaNDM-1 gene of A.baumannii was amplified by PCR and cloned into plasmid pMAL-p2X to construct recombinant plasmid pMAL-p2X ∶ ∶ NDM-1,based on which the bioinformatics of NDM-1 gene and protein were analyzed.The constructed recombinant plasmid was transformed to E.coli Tb1 and induced with IPTG,and the expressed recombinant protein MBP-NDM-1 was purified by affinity chromatography.Results Restriction analysis and sequencing proved that recombinant plasmid pMAL-p2X ∶ ∶ NDM-1 was constructed correctly.Bioinformatic analysis showed that the relative molecular mass and isoelectric point of NDM-1 were 28 487.86 and 5.89 respectively,while the signal peptide sequence was located in amino acids 1 ～ 28,and NDM-1 was low homologous to the metal β lactamase of other subtypes.The expressed recombinant protein,with a relative molecular mass of about 73 000,mainly existed in a soluble form,and reached a protein concentration of 1.72 mg / ml after purification.Conclusion Soluble recombinant protein MBP-NDM-1 was expressed and purified,which laid a foundation of further study on activity,function and inhibitor of NDM-1." @default.
- W2356390496 created "2016-06-24" @default.
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- W2356390496 date "2012-01-01" @default.
- W2356390496 modified "2023-09-28" @default.
- W2356390496 title "Cloning,expression and bioinformatics of NDM-1 gene" @default.
- W2356390496 hasPublicationYear "2012" @default.
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