人免疫球蛋白受体FcγRIIa的基因克隆及在K562细胞的表达  

CLONING HUMAN FcγRIIa AND EXPRESSION ON K562 CELLS

作　　者：田芳[1] 刘丹[1] 胡茂志[1] 龚卫娟[1] 季明春[1] 

机构地区：[1]扬州大学医学院,江苏扬州225001

出　　处：《实用临床医药杂志》2006年第2期36-39,共4页Journal of Clinical Medicine in Practice

基　　金：江苏省自然科学基金(BK2004404);江苏省高校自然科学基金(04KJB320162)

摘　　要：目的将外源人免疫球蛋白IgGFc段Ⅱ型受体CD32a分子表达于K562细胞表面,为构建人工抗原递呈细胞提供重要前提。方法自U937细胞RT-PCR得到CD32a的cDNA基因,与T载体连接后亚克隆入表达载体pcDNA3.1(+)。经测序后利用脂质体介导的转染将CD32a分子表达在K562细胞的表面。结果构建的T载体测序后,Genebank比对证实为CD32a的cDNA分子,免疫荧光和流式细胞仪结果均说明CD32a分子在K562细胞得到表达(96.9%)。结论获得的人工构建的表达人免疫球蛋白IgGFc受体的K562细胞,完成人工抗原递呈细胞制备的首要步骤。Objective In order to construct artificial antigen presenting cells, the exogenous receptor bound to the fragment IgG Fc were expressed on K562 cells. Methods The total RNA from U937 cell lines were isolated at first, reversed transcript reaction and PCR were used by oligodT primer and CD32a pair primers respectively, cDNA sequences were cloned into T vector. After sequencing, the CD32a were cloned into expressing vector pCDNA3.1 （ ＋ ）. Finally we expressed the CD32a molecules on K562 cells with the help of liposome. Results The results of sequencing from ABI 377 sequencer and blasting in the genebank show that we have got a cDNA sequence of CD32a. CD32a molecules have been expressed on K562 cells by the conformation of fluorescence microscope and flow cytometry. Conclusions We have got an artificial cell line from K562 cells that expresses the receptor of IgG Fc, which lays a solid foundation for constructing the artificial APCs in future.

关 键 词：CD32a K562细胞 表达 

分 类 号：R446[医药卫生—诊断学]