Title: Prokaryotic Expression Vector Construction, Expression and Purification of Human β-Actin Protein
Abstract: Objective:To construct the prokaryotic expression vector of human β-actin gene and to express and purify GST-β-actin fusion protein.Methods:Human β-actin coding region was amplified from human mammary gland cDNA by PCR,and was inserted into the prokaryotic expression vector of pGEX-KG.The recombinant plasmid GST-β-actin was transformed into E.coli Rossate.The expressed product was purified by GST-Sepharose 4B beads and identified by SDS-PAGE and Western blot analysis.Results:The DNA fragment of about 1100 bp was successfully amplified by PCR,and inserted into vector of pGEX- KG correctly.The results of double digestion and sequencing indicated that the GST-β-actin recombinant plasmid was successfully obtained.The GST-β-actin fusion protein of about M,40 000 was successfully induced,and identified by SDS-PAGE and Western blot analysis.Conclusion:The prokaryotic expression vector of GST- β-actin was constructed successfully,which lay the foundation for further research.
Publication Year: 2015
Publication Date: 2015-01-01
Language: en
Type: article
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