稳定表达人ASB12的C2C12细胞系的建立及鉴定  

Establishment and Identification of a Stable Human ASB12-Expressed C2C12 Cell Line

作　　者：文斗斗[1] 周军媚[2,3] 赵明一[2] 胡维新[1] 吴秀山[2] 王跃群[2] 

机构地区：[1]中南大学生物科学与技术学院,中国湖南长沙410013 [2]湖南师范大学,心脏发育研究中心,发育生物学教育部重点实验室,中国湖南长沙410081 [3]南华大学药学与生命科学学院,中国湖南衡阳421001

出　　处：《生命科学研究》2012年第4期283-286,共4页Life Science Research

基　　金：湖南省自然科学基金资助项目(11JJ3028);国家自然科学基金资助项目(81000035;31071999;30871340)

摘　　要：ASB12(homo sapiens ankyrin repeat and SOCS box containing 12)蛋白含有5个ANK(ankyrin repeat sequence)序列和一个保守的SOCS(suppressor of cytokine signaling)盒结构域,是ASBs(human ankyrin repeat andSOCS box containing protein family,ASB family)家族的成员.人类ASB12基因在成体心肌和骨骼肌组织中特异表达,是成肌分化的候选基因.利用阳离子聚合物转染技术将重组表达质粒pCMV-tag2B-ASB12转染小鼠骨骼肌细胞系C2C12细胞,通过G418筛选、免疫荧光检测、RT-PCR分析、Western blotting检测建立了稳定表达ASB12的细胞系C2C12-ASB12,为研究ASB12在骨骼肌发育及其相关功能提供有用的细胞研究模型.The human ASB12（Homo sapiens ankyrin repeat and SOCS box containing 12） protein contains five ANK（ankyrin repeat sequence） domains and a SOCS（suppressor of cytokine signaling） box domain,be-longing to the ASBs family.It was reported that ASB12 especially expressed in skeletal and cardiac muscles of adult tissues,which suggested that ASB12 closely associated with skeleton muscle development.To con-struct a stable ASB12-expressed C2C12 cell line,the fusion expression plasmid pCMV-tag2B-ASB12,which was identified by enzyme digestion and sequencing analysis,was transfected into C2C12 cell by cationic polymer.After screening culture by G418,the expression of ASB12 was detected by immunofluorescence,RT-PCR and Western-blotting.The C2C12 cell line that expressing ASB12 stably was established successfully,which provide a cell model for studying the molecular function of ASB12 in skeleton muscle development.

关 键 词：ASB12 ANK结构域 SOCS box结构域 C2C12稳定细胞系 

分 类 号：Q28[生物学—细胞生物学]