PH STABILIZING CHO/DHRF/ANTI-LDH CELL LINE REDUCING LACTATE PRODUCTION BY USING ANTISENSE MRNA TECHNIQUES

摘要

PURPOSE: A pH stabilizing CHO/dhrf/anti-ldh cell line reducing lactate production by using antisense mRNA techniques is provided, thereby mass-producing proteins by inhibiting acidification due to lactate production. CONSTITUTION: An antisense nucleotide anti-ldh inhibits the production of lactate dehydrogenase(LDH) of Chinese Hamster Ovary(CHO) and has the nucleotide sequence complementary with CHO dhrf-LDH-A cDNA of SEQ ID NO: 1. A fusion gene dhfr/anti-ldh is prepared by fusion of the antisense nucleotide anti-ldh with DHFR gene (ATCC 37146). An animal cell expression vector contains the fusion gene dhfr/anti-ldh. The pH stabilizing CHO/dhrf/anti-ldh cell line is prepared by transforming CHO dhrf- cell line with the animal cell expression vector, and amplifying the fusion gene within the transformed CHO dhrf- cell line using methotrexate(MTX).