Expression vector containing urea cycle enzyme gene, transformant thereof, and the use of transformant for protein overexpression

摘要

PURPOSE: Provided are an expression vector including a gene of urea cycle in a living body, a transformant transformed with the vector and its use for the over-expression of proteins, thereby reducing the accumulation of ammonia in a culture medium and increasing the growth rate of cells to obtain proteins in a high yield. CONSTITUTION: The expression vectors are as follows: pCIneo-CPS45(KCTC 0846BP) including rat CPS I gene; pREP4-OTC1(KCTC 0847BP) including rat OTC gene; pc3.1/Zeo(+)-AS(KCTC 10014BP) including rat AS gene; pc3.1/Zeo(+)-AL(KCTC 10013BP) including human AL gene; and pc3.1/Zeo(+)-Arg(KCTC 10012BP) including rat Arg gene. Animal cells are transformed with them and express CPS I, OTC, AS, AL and Arg genes. The animal cells are selected from the group consisting of CHO(Chinese Hamster Ovary) cell; COS-7(Monkey, African green) cell; NIH/3T3(Mouse fibroblast) cell; BHK-21(Baby Hamster Kidney, Syrian golden) cell; Hela(Human carcinoma) cell; Vero(Monkey kidney, African green) cell; and C127 (Mouse) cell. The transformants used for the over-expression of proteins are as follows: CHO-OTC1-A19(CHO/dhfr¬-)(KCTC 0845BP); CHO-AS-16(CHO/dhfr¬-)(KCTC 10031BP), CHO-AL-19(CHO/dhfr¬-)(KCTC 10015BP); and CHO-Arg-11(CHO/dhfr¬-)(KCTC 10030BP).