Abstract: Objective To study the expression of rhIFN-α2b gene in pET system using constructed vector.Methods Amplify human IFN-α2b gene by PCR using synthesized primers, and change the 4 rare codons into biased ones of E. coli by introducing site-directed mutagenesis. In this way, all the base sequences encoding the first 16 amino acids after the start codon were changed into E. coli biased, however, the ami-no acid sequence identical to natural IFN-α2b was unchanged. Subclone the PCR product into pET28a vector. Transform the recombinant plasmid pET28a- IFN-α2b to competent E. coli strain BL21(DE3) and express under induction of EPTG. The expressed product was primarily purified and detected for IFN-α2b activity. Results The expressed rhIFN-α2b existed in the form of soluble protein, and showed high biological activity compared with that expressed in pBV889 system. Conclusion pET system can be used for the expression of rhIFN-α2b protein with higher biological activity.
Publication Year: 2003
Publication Date: 2003-01-01
Language: en
Type: article
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