Construction and Identification of the Recombinant Prokaryotic Expression Plasmid pQE30-bgln

A recombinant plasmid pQE30-bgln was constructed for obtaining the BGLN protein expressed in E. coil so as to develop the efficiency of extracting resveratrol from plants. Touch-down PCR was used to amplify the bgln gene from plasmid pUCP67 in which bgln was cloned. A new plasmid pQE30-bgln, was then constructed by inserting the amplified bgln gene into pQE30, a prokaryotic expression vector. Restriction analysis and sequencing were used to confirm the structure of pQE30-bgln. Results showed that a DNA fragment in size of 2.2 kb was amplified. Furthermore, restriction analysis showed that the amplified gene was inserted into pQE30 correctly and the size of recombinant plasmid is abont 5.6 kb. The forepart interface between the vector and the insert was sequenced by TaKaRa company, and it was confirmed that the insert was not out of frame. In conclusion, the plasmid pQE30-bgln is constructed successfully.