摘要
目的 建立体细胞GPA突变频率的检测方法 ,研究环境基因毒素如电离辐射等对人体的损伤。方法 用自制 3株对M和N型血型糖蛋白A(GlycophorinA ,GPA)的特异单克隆抗体 :3G4(抗M ) ,6A8(抗N)和 3C5(抗MN) ,进行荧光素和生物素标记 ,与甲醛和 /或二甲亚胺固定的人外周血红细胞结合 ,在流式细胞上以荧光强度分选出正常和变异红细胞 ,分析MN型个体红细胞突变性质 (即MN→MO、MM、NO、NN) ,计算突变频率。结果 建立了GPA分析法 1Wa、1Wb和 2Wa法 ,1Wa、1Wb法可分别检测MN血型个体红细胞的MO、MM突变细胞群和NO、NN突变细胞群 ,2Wa法可将MN型个体红细胞分选出MO、MM、NO、NN型 4种突变细胞 ,并确定变异细胞频率。
Objective\ To study the damage to human body caused by environmental radiation,and supervise the somatic mutations. Methods Three monoclonal antibodies specific to M\|type(3G4),N\|type(6A8), and MN\|type (3C5) of glycophorin A,respectively,were prepared.Fluorescence or biotin conjugated antibodies were bound specifically to formalin and/or dimethyl suberimidate fixed erythrocytes.M,MN,and N type cells were divided by cytometry to demonstrate the erythrocyte mutation characteristics (MN→MO,MM,NO,NN) and give out the variant frequency. Results 1Wa,1Wb and 2Wa methods of GPA assay were developed.Erythrocytes of MN type individuals could be separated to normal and single locus variant groups by 1W methods and they could be sorted as normal (MN),single gene deletion mutants (MO,NO),homozygous mutants (MM,NN) cell groups by 2Wa method. Conclusion The assay is applicable to evaluationg the frequency of variant erythrocytes from human somatic mutation.\;
出处
《中华放射医学与防护杂志》
CAS
CSCD
北大核心
2000年第3期164-167,共4页
Chinese Journal of Radiological Medicine and Protection
基金
解放军总后"九.五"课题基金资助项目! ( 96 0 50 0 2 )
