[一种候选人类Era结合蛋白A19的表达及其多克隆抗体的制备]
[Expression of a candidate human Era binding protein A19 and the preparation of its polyclonal antibody].
作者信息
Li Yun-feng, Wu Yuan-ming, Chen Su-min, Chen Nan-chun, Huang Yong, Zhang Xiao-nan
机构信息
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 Jul;21(4):459-62.
AIM
To express a candidate hEra binding protein A19 in Escherichia coli and to prepare anti-A19 antibody.
METHODS
A19 gene was amplified by PCR from the plasmid containing A19 gene and was cloned into the expression vector pGEX-4T3 which was then transformed into E.coli. The A19 protein was expressed under IPTG induction. Antiserum was prepared by immunizing rabbits with the expressed A19 protein. The titer and specificity of polyclonal antibody were detected by Western blot.
RESULTS
The expressed A19 accounted for about 30.2% of total bacterial protein. The titer of the antiserum was about 1:4 000. Western blot analysis indicated that the antiserum had high specificity.
CONCLUSION
A19 fusion protein was highly expressed. The specific anti-A19 antiserum was prepared successfully.
目的
在大肠杆菌中表达候选hEra结合蛋白A19并制备抗A19抗体。
方法
通过PCR从含有A19基因的质粒中扩增A19基因，并将其克隆到表达载体pGEX-4T3中，然后将其转化到大肠杆菌中。A19蛋白在IPTG诱导下表达。用表达的A19蛋白免疫兔子制备抗血清。通过Western印迹检测多克隆抗体的效价和特异性。
结果
表达的A19约占细菌总蛋白的30.2%。抗血清效价约为1:4000。Western印迹分析表明抗血清具有高特异性。
结论
A19融合蛋白高表达。成功制备了特异性抗A19抗血清。
Zotero 插件