乙酰肝素酶可变剪接体Splice 5在CHO细胞中的稳定转染及其亚细胞定位研究  

Subcellular localization of heparanase variant Splice 5 in stably transfected CHO cells

作　　者：顾懿[1] 高红伟[2] 李素波[2] 金泗虎[2] 鲍国强[2] 檀英霞[2] 王颖丽[2] 季守平[2] 裴雪涛[1,2] 宫锋[2] 

机构地区：[1]广西医科大学,南宁530021 [2]军事医学科学院野战输血研究所,北京100850

出　　处：《军事医学》2013年第7期514-516,共3页Military Medical Sciences

摘　　要：目的建立乙酰肝素酶可变剪接体Splice 5稳定转染中国仓鼠卵巢(CHO)细胞株,并进行亚细胞定位分析。方法将表达乙酰肝素酶全长的质粒和乙酰肝素酶可变剪接体Splice 5质粒转染入CHO细胞,经G418进行筛选后,挑取单克隆进行培养,RT-PCR检测筛选克隆。用激光共聚焦显微镜测定其亚细胞定位。结果成功获得乙酰肝素酶可变剪接体Splice 5稳定转染株,并研究了其亚细胞定位情况。野生型乙酰肝素酶以颗粒状分布于细胞质中,Splice 5以弥散状分布于细胞质中,且在内质网、高尔基体、溶酶体、细胞膜均有分布。结论 Splice 5和野生型乙酰肝素酶在CHO细胞中分布形式和定位的不同,提示Splice 5可能具有其他的生物学功能。Objective To establish spliced variant of human heparanase（ Splice 5） stably transfected Chinese hamster ovary（CHO） cell lines and to investigate the subcellular localization of Splice 5. Methods The plasmid containing Splice 5 was transfected into CHO ceils which were selected by G418. Stable cell lines were identified by RT-PCR. The subcellu- lar localization of heparanse and its variant Splice 5 was observed by laser confocal microscope. Results CHO cell lines stably expressed Splice 5 were successfully established. Wild-type heparanase was predominantly distributed via granular pattern in the cytoplasm, while Splice 5 was dispersed and also distributed in the endoplasmic reticulum, Golgi apparatus, lysosomes and cell membrane. Conclusion Splice 5 shows different localization with wild type heparanase and may perform other functions in CHO cells.

关 键 词：乙酰肝素酶 可变剪接体 转染 亚细胞定位 

分 类 号：R341[医药卫生—基础医学]