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在DNA转染细胞中建立血型ABO特异性转移酶活性的简易检测系统。

Establishment of a simple detection system for blood group ABO-specific transferase activity in DNA-transfected cells.

作者信息

Nishida Chiyomi, Nishiyama Chiharu, Satoh Koichi, Hara Mutsuko, Itoh Yukio, Ogawa Hideoki, Okumura Ko

机构信息

Department of Forensic Medicine, Juntendo University School of Medicine, Tokyo, Japan.

出版信息

Leg Med (Tokyo). 2010 Jul;12(4):172-6. doi: 10.1016/j.legalmed.2010.03.005. Epub 2010 May 14.

Abstract

A/B-transferase is a glycosyltransferase that transfers a sugar substrate onto H-antigen resulting in the synthesis of glycoproteins and glycolipids termed A/B-antigens. The ABO blood group (ABO) gene encoding A/B-transferase possesses numerous polymorphisms affecting the specificity and/or activity of the enzyme. The relationship between genotype and phenotype is very complicated, except for those of some critical polymorphisms. In order to establish a system for evaluating the effect of each polymorphism on the transferase function, an A- or B-transferase cDNA expressing vector was introduced into HeLa cells, a cell line that do not possess endogenous A/B-transferase activity. We successfully detected substrate-specific transferase activity in the cells and in the culture medium. Furthermore, in three different assays, each corresponding A- or B-antigen was detected in the transfectants with high sensitivity. Accordingly, the present study demonstrates a possibility that A/B-transferase variants may be characterized by using this method.

摘要

A/B-转移酶是一种糖基转移酶,它将糖底物转移到H抗原上,从而合成称为A/B抗原的糖蛋白和糖脂。编码A/B-转移酶的ABO血型(ABO)基因具有许多影响该酶特异性和/或活性的多态性。除了一些关键多态性的情况外,基因型和表型之间的关系非常复杂。为了建立一个评估每种多态性对转移酶功能影响的系统,将A或B转移酶cDNA表达载体导入HeLa细胞,这是一种不具有内源性A/B转移酶活性的细胞系。我们成功地在细胞和培养基中检测到了底物特异性转移酶活性。此外,在三种不同的检测方法中,在转染子中以高灵敏度检测到了各自相应的A或B抗原。因此,本研究证明了使用这种方法表征A/B转移酶变体的可能性。

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