摘要
利用RT-PCR扩增法获取山羊Wnt10b基因序列并将该目的片段连接到原核表达载体p ET32a,转化E.coli表达菌BL21(DE3),构建了Wnt10b/p ET32a(+)-BL21(DE3)工程菌,以IPTG诱导表达Wnt10b融合蛋白,Ni柱亲和层析纯化蛋白,将获得抗原免疫新西兰大白兔,获得了1:243000效价的山羊Wnt10b基因的多克隆抗体,为后续山羊Wnt10b基因功能研究提供参考.
The WntlOb CDS, amplified by reverse transcription-polymerase chain reaction（ RT-PCR）, was inserted into pET32a vector,and then transformed into E. coli BL21 （DE3） to construct the WntlOb/pET32a （ ＋ ） - BI221 （DE3） bioengineering strain,and finally WntlOb fusion protein was induced by IPTG. The WntlOb protein was purified by Ni column affinity chromatography, and further used for the immunization of New Zealand white rabbits. The polyclonal antibody of goat WntlOb with titer of 1：243000 was obtained. The above work provided relevant material for further study of goat WntlOb functions.
作者
马洁琼
林亚秋
朱江江
黄凯
王永
MA Jie-qiong;LING Ya-qiu;ZHU Jiang-jiang;HUANG Kai;WANG Yong(School of Life Science and Technology, Southwest Minzu University, Chengdu 610041, P. R. C.;Key Laboratory of Sichuan Province for Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation, Southwest Minzu University, Chengdu 610041, P. R. C.)
出处
《西南民族大学学报（自然科学版）》
CAS
2018年第3期250-253,共4页
Journal of Southwest Minzu University(Natural Science Edition)
基金
四川省"十三五"畜禽育种攻关项目(2016NYZ0045)
