Title: Cloning and expression of recombinant onconase in Escherichia coli
Abstract: Based on the amino acid sequence of onconase and the preferred codons of the bacterium Escherichia coli,the target gene was synthesized by PCR-driven overlap extension of about 350 bp and linked into the pGEM-T vector.After sequencing,the gene was subcloned into the expression vector pET-22b(+) to obtain the recombinant expression vector,which was transformed into Rosetta.Positive colonies were confirmed by PCR and sequencing.In order to check the gene expression in Rosetta,tricine-SDS-PAGE was performed on the target protein.The results showed that the gene was expressed successfully with a content of 48.8%.Moreover,the proposed primary structure of the expressed protein was shown to be correct,with a coverage of 35%,by characterization by LC-ESI-MS/MS with the LCQ DECA XP Plus system.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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