Construction of secretory expression system suitable to express glucagon under the control of PL promoter.
It was reported that PL promoter and alkaline phosphatase (phoA) signal peptide were used to construct secretory expression plasmid suitable to express glucagon and [Des-His1] glucagon in E. coli BL21 herein. Expression studies showed these two peptides could be expressed and secreted into the culture medium. The expression yield of recombinant glucagon reached 3.46 mg/L/OD600 unit of cells in shake flask. The yield of [Des-His1] glucagon was found to be higher than that of glucagon. In addition, some factors involved in secretion were studied too.
Institute of Biochemistry and Cell Biology, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, People's Republic of China.
Chongwei
C
Wen
Renbao
R
Gan
Shangquan
S
Zhu
D000469:Alkaline Phosphatase / Q000235:genetics; D000595:Amino Acid Sequence; D015153:Blotting, Western; D003470:Culture Media / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D005934:Glucagon / Q000031:analogs & derivatives / Q000235:genetics* / Q000378:metabolism*; D002479:Inclusion Bodies / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000235:genetics; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013696:Temperature; D014169:Transformation, Bacterial
D003470:Culture Media; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D005934:Glucagon; D000469:Alkaline Phosphatase