Expression and purification of delta sleep-inducing peptide in Escherichia coli
- Authors
- Oh, Kwang-Seok; Na, Do-Kyun; Kweon, Mee-Hyang; Sung, Ha-Chin
- Issue Date
- Aug-2003
- Publisher
- KOREAN SOC MICROBIOLOGY & BIOTECHNOLOGY
- Keywords
- DSIP; expression; peptide production; labeling of small peptide
- Citation
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY, v.13, no.4, pp 620 - 623
- Pages
- 4
- Journal Title
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
- Volume
- 13
- Number
- 4
- Start Page
- 620
- End Page
- 623
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/43528
- ISSN
- 1017-7825
1738-8872
- Abstract
- The delta sleep-inducing peptides (DSIP, Trp-Ala-Gly-Gly-Asp-Ala-Ser-Gly-Glu) is an important regulatory hormone, controlling hypothalamus and pituitary functions. In the Current study, an expression system was designed for the rapid and economic expression of recombinant DSIP for biophysical studies. Artificially synthesized oligonucleotides encoding DSIP were cloned into a pGEX-KG vector and expressed in E. coli (BL21). The recombinant GST-DSIP was then readily purified using a GST affinity Column. To obtain intact DSIP from the GST-DSIP, thrombin cleavage and a CNBr reaction were successively carried out. The DSIP in the CNBr reaction mixture was subjected to RP-HPLC purification to yield 1.2 mg DSIP from a 1 liter Culture of E. coli. Identification of the DSIP was performed using MALDI-MS and an amino acid composition analysis.
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