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| 蛋白激酶Cα-EGFP及其突变体真核表达载体的构建和在C2C12细胞中的表达 | |
| 引用本文： | 陈瑞华,张耕,陆纲,鹿培源,贾弘禔.蛋白激酶Cα-EGFP及其突变体真核表达载体的构建和在C2C12细胞中的表达[J].河北北方学院学报(医学版),2001,18(2):5-7. |
| 作者姓名： | 陈瑞华 张耕 陆纲 鹿培源 贾弘禔 |
| 作者单位： | 1. 张家口医学院生物教研室,075000 2. 张家口医学院组胚教研室,075000 3. 北京大学医学部生物化学与分子生物学系,100083 |
| 摘 要： | 目的研究PKCα结构与转位的关系.方法用PCR的方法构建了4个PKCα突变体,在DNA连接酶的作用下与pEGFP-N1结合后转移至质粒中,构建PKCα突变体-EGFP真核表达载体.观察它们在C2C12细胞中的表达及转位情况.结果pPKCα-EGFP-N1转染C2C12肌细胞24h后,细胞内表达的PKCα-GFP融合蛋白主要定位于细胞浆,细胞核中未见分布.pPKCα-βI-EGFP-N1、pPKCβI-α-EGFP转染C2C12细胞24h的表达产物集中在胞浆.pPKCα(-)-EGFP-N1、pPKCα(m)-EGFP-N1转染C2C12细胞24h的定位与野生型明显不同.结论PKCα的调节区(C1V1C2)尤其是RACK结合区(C2)、PKCα的催化区(C3V4C4)尤其是ATP结合位点与PKCα的转位密切相关. |
| 关 键 词： | 蛋白激酶C PKCα突变体 表达 转位 绿色荧光蛋白 |
| 文章编号： | 1005-8796(2001)02-005-03 |
Construction of eucaryotk expression vectors of protein Kinase C a- EIGFP and it's mutants and their expression in C2C12 cells |
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| Chen Rui hua,Zhang Geng,Lu Gang,et al.Construction of eucaryotk expression vectors of protein Kinase C a- EIGFP and it''''s mutants and their expression in C2C12 cells[J].Journal of Hebei North University:Medical Edition,2001,18(2):5-7. | |
| Authors: | Chen Rui hua Zhang Geng Lu Gang |
| Institution: | Chen Rui hua,Zhang Geng,Lu Gang,et al Department. of Biology,ZhangJiaKou Medical College,zhangjiakou 075000 China |
| Abstract: | Objective To explore the relationship between the structure of protein kinase C a(PKCa) and it's translocation in C2C12 cells. Methods:Four mutants of PKCa developed by PCR method were combined with pEGFP - N 1by DNA ligase and transferred into plasmids to construct eucaryotic expression vectors of PKC mutant - EGFP. Their expression and translocation in C2C12 cells were observed. Results: The PKCa - GFP fusbn proteins were mainly anchored in cytoplasm of C2C12 cells, The location of pPKCa( - ) - EGFP - Ml or pKCa (m) -EGFP-N1 in C2C12 cells after transfection for 24hs was obviously different form that of wild types. Conclusion: The regulatory site(C1V1C2) of PKCa, especially combining site(C2) of RACK is related to it's translocation, so is the ctalytic site(C3V4C4) of PKC, especially combining site with ATP. |
| Keywords: | Protein Kinase Ca PKCa mutant - EGFP Expression Translocation |
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