[Expression and identification of PEBP-like gene from Saussurea involucrate Kar.er kir in Escherichia coli].
This assay was designed to construct the prokaryotic expression vector, investigate the expression of PEBP-like in Escherichia coli and purify its product. The PEBP gene was inserted into the vector pET30a (+). The recombinant vector was transferred into E. coli BL21 (DE3)and induced the expression of protein by low concentration of IPTG and low temperature overnight. After purification, the supernatants were analyzed by SDS-PAGE and the results were identified by Western blotting. After IPTG induction, a new anticipating fusion protein of 28 kD appeared as an expected size, and its product was 26.8% in total protein, the fusion protein was positive by Western blotting. The prokaryotic expression system of PEBP-like is successfully constructed. It lays the foundation for the further application study on the antifreeze characters of the PEBP.
Wang
Zhifang
Z
Institute of Microbiology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China.
Zhang
Xuejun
X
Wang
Bo
B
Zhou
Xiaoyun
X
Ai
Xiulian
X
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D049488:Phosphatidylethanolamine Binding Protein / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D030441:Saussurea / Q000235:genetics*
D049488:Phosphatidylethanolamine Binding Protein; D010940:Plant Proteins; D011994:Recombinant Proteins