正义和反义表达grp75基因的哺乳动物细胞克隆的建立  

Establishment of mammalian cell lines with sense and antisense expression of grp75 gene.

作　　者：左伋[1] 荣琦[1] 郑东航[1] 夏蓓莉[1] 陈秀珍[1] 

机构地区：[1]上海医科大学医学遗传学研究室,上海200032

出　　处：《中国优生与遗传杂志》1998年第1期1-2,7,共3页Chinese Journal of Birth Health & Heredity

基　　金：国家自然科学基金资助(39670239);美国中华医学基金会资助项目(96-635)

摘　　要：为了进一步研究葡萄糖调节蛋白(grp75)基因及其产物的生理功能和它在疾病发生发展过程中的作用,以 peDNA3为载体构建了正义(向)和反义(向)的 grp75 cDNA 真核细胞表达载体系统,并对转染哺乳动物(PC12)细胞系后所筛选得到的阳性克隆进行免疫印迹分析;结果表明转染了正向grp75 cDNA 表达系统的细胞克隆在原有 GRP75表达的基础上表达量显著增加;而转染了反向 grp75cDNA 表达乐统的细胞克隆中 GRP75的表达水平明显下降,说明所构建的正向及反向 grp 75cDNA 表达系统在哺乳动物细胞系中得到了有效的表达,显示所构建的表达载体及阳性细胞系将可用于 grp75基因的生理和病理研究中。In order to obtain mammalian cell lines with sense and antisense expression of grp75 cDNA which may be suitable for the investigation of physiological functions and pathological effects of grp75,grp75F and grp75B expression vectors were ligated by means of recombinant DNA techniques and were proved by enzymatic analysis and se- quencing.The constructed vectors were then transfected into mammalian cell (PC12) line,in which the expressed products were deteeted by im- munoblotting.The result revealed that grp75F in- creased the level of GRP75 dramatically(sense ex- pression);while grp75B decreased the amount of GRP75 significantly (antisense expression).The woek suggest that the mammalian cells with sense and antisense expression of grp75 gene are estab- lished and all of these cells can be used in the fur- ther studies of grp75 gene and its peptide.

关 键 词：GRP75 基因表达 载体 哺乳动物 细胞系 克隆 

分 类 号：Q343[生物学—遗传学]