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KMID : 0545120030130040620
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Journal of Microbiology and Biotechnology
2003 Volume.13 No. 4 p.620 ~ p.623
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Expression and Purification of Delta Sleep-Inducing Peptide in Escherichia coli
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OH, KWANG-SEOK
NA, DO-KYUN/KWEON, MEE-HYANG/SUNG, HA-CHIN
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Abstract
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The delta sleep-inducing pcptides (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 recornbinant DSIP for biophysical studies. Artificially synthesized oligonucleotides encoding DSIP were cloned into a pGEX-UG 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 lo RE-HPLC purification to yield 1.2㎎ DSlP from a 1 liter culture of E. culi. Identification of the DSlP was performed using MALDT-MS and an amino acid composition analysis.
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KEYWORD
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