八肋游仆虫第一类肽链释放因子eRF1a多克隆抗体的制备、纯化及鉴定  

Preparation,Purification and Identification of Polyclonal Antibody against Class I Release Factor eRF1a from Euplotes octocarinatus

作　　者：梁爱华[1] 宋莉[1] 张志云[1] 

机构地区：[1]山西大学生物技术研究所化学生物学与分子工程教育部重点实验室,山西太原030006

出　　处：《山西大学学报（自然科学版）》2007年第2期257-260,共4页Journal of Shanxi University(Natural Science Edition)

基　　金：国家自然科学基金(3067028230470239);山西省自然科学基金(2004107920051065)

摘　　要：将八肋游仆虫第一类肽链释放因子基因eRF1a克隆入表达载体pRSETc,并在大肠杆菌Bl21(DE3)中进行诱导表达.表达形成的包含体经切胶回收,然后免疫大鼠制备抗血清.所获得的抗血清用Amersham Biosciences抗体纯化试剂盒进行纯化,SDS-PAGE电泳分析表明纯化后的抗体纯度很高.Western blotting印迹和ELISA鉴定结果表明抗体特异性好,效价为1∶5000.Euplotes octocarinatus eRF1a gene was cloned into the expression vector pRSETc. The obtained plasmid pRSETC-eRF1a was transformed into E. coli BL21 （DE3）,and the fusion protein His6-eRF1a was expressed. The wistar rat was injected with purified His6-eRF1a inclusion bodies to induce the immunoreactions. The antiserum was purified with Mab Trap Kit and detected by SDS-PAGE,ELISA and Western blotting. The results showed that the polyclonal antibody had high purity and specificity. The antibody titer detected by ELISA reached to 1 ： 5 000.

关 键 词：八肋游仆虫 肽链释放因子 eRF1a 多克隆抗体 

分 类 号：Q78[生物学—分子生物学]