斑马鱼charon基因的克隆、抗体制备及分析  

Cloning and Polyclonal Antibody Preparation of charon Gene in Zebrafish

作　　者：吴坚[1] 王玲[1] 戴悦[1] 吴秀山[1] 王跃群[1] 

机构地区：[1]湖南师范大学蛋白质化学及鱼类发育生物学教育部重点实验室心脏发育研究中心,湖南长沙410081

出　　处：《激光生物学报》2011年第3期347-352,共6页Acta Laser Biology Sinica

基　　金：国家自然科学基金项目(31071999;30871340;30930054);国家重点基础研究发展计划资助顺目(2005 CB522505)

摘　　要：斑马鱼charon基因位于1号染色体上,其成熟肽编码区含有731bp,编码243个氨基酸。charon基因编码Cerberus/Dan家族的一种分泌因子,它与心脏的左右不对称发育有关。为进一步研究charon在心脏左右不对称发育中的功能,以斑马鱼为动物模型,利用RT-PCR的方法成功克隆了斑马鱼charon基因片段,并将其进行原核表达,制备了charon多克隆抗体。通过western blotting对抗体进行分析,结果表明:实验获得了效价较高、特异性较好的charon多克隆抗体,而且charon在斑马鱼中表达较为广泛,在心脏、肌肉、眼睛等组织表达尤其明显。The charon gene in zebrafish is located on chromosome 1 , which coding region has 731 bp and encodes 243 amino acids. Charon encodes a Cerberus/Dan family secreted protein, which is related in left/right asymmetric heart development. Zebrafish gene charon has homologous genes in humans, mice and rats. In order to study the function of charon in left/right asymmetric heart development, a fragment of zebrafish gene charon was cloned by RT- PCR technique, then induced to express charon fushion protein with prokaryotie expression. We successfully prepared charon polyclonal antibodies. The antibody is assayed by western blotting and the result showed that the high quality multi-clonal antibody was obtained and charon is expressed extensively in the zebrafish, particularly evident in the heart, muscle and eye.

关 键 词：CHARON 融合蛋白 斑马鱼 原核表达 多克隆抗体 

分 类 号：Q786[生物学—分子生物学]