S Tao1, Y He, Y Ying, X Hong, X Xu, F Zhu, H Lv, L Yan. 1. Blood Center of Zhejiang Province, Wulin Road 345, Hangzhou, Zhejiang Province 310006, People's Republic of China.
Abstract
PURPOSE OF THE STUDY: The C35T mutant α-(1,2)-fucosyltransferase (FUT1) gene had been reported related to the para-Bombay phenotype, but recently, our laboratory found that the C35T was a polymorphism in the Chinese population. This study aims at further clarifying the property of C35T mutant FUT1. MATERIAL AND METHODS: The mutant C35T FUT1 gene was cloned into expression vector in vitro, the mRNA expression was analyzed by real-time quantitative polymerase chain reaction (PCR), and the mutant enzyme's activity was determined in vitro. RESULTS: The results showed that the frequencies of 35C and 35T alleles were 0.735 and 0.265, respectively. The FUT1 mRNA level of transfected cells with C35T recombination vector showed 99.85% of the wild-type FUT1 transfected cells. The enzyme relative activity of transfected cell lysates with C35T FUT1 recombination vector was 79.45% compared with that of the wild-type FUT1 transfected cell lysates. The K(m)(phenyl-gal) value of enzyme encoded by C35T allele was 0.5 times higher than that of the enzyme encoded by FUT1 wild-type allele. CONCLUSION: These results suggested that FUT1 C35T was a polymorphism in the Chinese population and did not affect its mRNA transcription, but could slightly decrease the activity of human α-(1,2)-fucosyltransferase in vitro.
PURPOSE OF THE STUDY: The C35T mutant α-(1,2)-fucosyltransferase (FUT1) gene had been reported related to the para-Bombay phenotype, but recently, our laboratory found that the C35T was a polymorphism in the Chinese population. This study aims at further clarifying the property of C35T mutant FUT1. MATERIAL AND METHODS: The mutant C35TFUT1 gene was cloned into expression vector in vitro, the mRNA expression was analyzed by real-time quantitative polymerase chain reaction (PCR), and the mutant enzyme's activity was determined in vitro. RESULTS: The results showed that the frequencies of 35C and 35T alleles were 0.735 and 0.265, respectively. The FUT1 mRNA level of transfected cells with C35T recombination vector showed 99.85% of the wild-type FUT1 transfected cells. The enzyme relative activity of transfected cell lysates with C35TFUT1 recombination vector was 79.45% compared with that of the wild-type FUT1 transfected cell lysates. The K(m)(phenyl-gal) value of enzyme encoded by C35T allele was 0.5 times higher than that of the enzyme encoded by FUT1 wild-type allele. CONCLUSION: These results suggested that FUT1C35T was a polymorphism in the Chinese population and did not affect its mRNA transcription, but could slightly decrease the activity of human α-(1,2)-fucosyltransferase in vitro.