Journal
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
Volume 106, Issue 2, Pages 168-173Publisher
SOC BIOSCIENCE BIOENGINEERING JAPAN
DOI: 10.1263/jbb.106.168
Keywords
Chinese hamster ovary cells; fucosylation; small interfering RNA; antithrombin III; GDP-fucose transporter
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The full-length cDNA of GDP-fucose transporter (GFT) was cloned from Chinese hamster ovary (CHO) 13D-35D cells. The predicted product consisted of 366 amino acids with 86% and 92% similarities to human GIFT and mouse GIFT, respectively. When synthetic small interfering RNA (siRNA) against CHO GFT was introduced into human antithrombin III (AT-III)-producing recombinant CHO cells, defucosylated AT-III increased by 30-40% after 3-d incubation. A GFT siRNA expression plasmid was also constructed to defucosylate recombinant AT-III. CHO cells, which were transfected with the siRNA expression plasmid, showed approximately reduction of Golgi-GDP fucose, resulting in 10-40% increase in defucosylated AT-III.
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