PROMOTER PORTION OF ALGINATE LYASE DERIVED FROM PSEUDOMONAS SP.

摘要

PURPOSE: A method for preparing an expression vector for highly expressing growth hormone using a promoter portion of alginate lyase derived from Pseudomonas sp. is provided, thereby rapidly and cheaply producing the growth hormone. CONSTITUTION: An expression vector pALN69 for highly expressing growth hormone in E. coli contains a promoter and the growth hormone of Gilthead seabream. A method for preparing the expression vector comprises the steps of: PCR amplifying cDNA of Gilthead seabream growth hormone in a vector pGSBGH3 using primers O-I, O-II and inserting SalI and HindIII recognizing sites to both ends of the amplified cDNA; ligating the cDNA fragment with a promoter portion of alginate lyase of Pseudomonas sp. using T4 ligase; PCR amplifying the ligated gene fragment using primers O-II, O-III; adding a restriction enzyme EcoRI recognizing site to the front portion of the amplified promoter; and ligating the amplified promoter with a cloning vector pUC19.