生命科学研究2013，Vol.17Issue(3)：196-199,4.
gcm蛋白的表达、纯化及多克隆抗体的制备
Expression, Purification of gcm Fusion Protein and Preparation of its Polyclonal Antibody
摘要
Abstract
gcm(glial cells missing) appears to function as a binary genetic switch for glia versus neurons.In loss-of-function gcm mutant alleles,presumptive glia become neurons; In gain-of-function gcm conditions,presumptive neurons become glia.Meanwhile,gcm control plasmatocyte development.Obtaining the gcm protein and it's antibody is the first step to detect the function of gcm in heart development.The cDNA fragment of part coding sequence of gcm was obtained by PCR amplification,then the fragment was cloned into PET-28a vector and identified with restriction enzyme and sequencing.The recombinant expression plasmid containing gcm gene was transformed into BL21 and fusion protein was induced by IPTG.The purified protein was obtained by treating the lysates with immobilized Glutathione Sepharose.The protein product was used to immune the New Zealand white rabbits to prepare antibody.The antibody titer and specification was identified by Western blot.We have successfully obtained GST-tag fusion protein of gcm and got high sensitivity and specificity anti-gcm polyclonal antibody,which lays a solid foundation for the further studies of gcm function.关键词
gcm/融合蛋白/多克隆抗体Key words
gcm/fusion protein/polyclonal antibody分类
生物科学引用本文复制引用
赵阳,彭佩莹,王静,周英,张义,唐超,任恋,彭夕洋,陈思行..gcm蛋白的表达、纯化及多克隆抗体的制备[J].生命科学研究,2013,17(3):196-199,4.基金项目
湖南省普通高校学科带头人培养计划项目(湘教通2008-315) （湘教通2008-315）
