Dong Ke, Chen Su-min, Ji Zong-ling, Zheng Yu, Chen Nan-chun
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 May;21(3):280-3.
To express and purify two alternative splicing mouse Era proteins and detect whether anti-human Era antibody can be used in the study of mouse Era proteins.
Two fusion protein expression vectors, pMAL-meraW and pMAL-meraS, were constructed, then the MBP-mEra proteins were expressed in E. coli. The target proteins were purified by amylose affinity chromatography. The specificity of rabbit anti-human Era antibody to the proteins was identified by Western blot.
The expressed MBP-mEraW and MBP-mEraS proteins constituted approximately 17% and 19% of the total bacterial proteins. The purity of the fused proteins was 67% and 61% respectively after amylose affinity chromatography. Rabbit anti-human Era antibody had high specificity to these two kinds of splicing mouse Era proteins.
Two fusion mera genes could be expressed in E. coli by using gene recombination technique. The high specificity of rabbit anti-human Era antibody to the two splicing mouse ERA proteins indicates that this antibody can be used to study the function of these two kinds of splicing mouse Era.
表达并纯化两种可变剪接的小鼠Era蛋白，检测抗人Era抗体是否可用于小鼠Era蛋白的研究。
构建两种融合蛋白表达载体pMAL-meraW和pMAL-meraS，然后在大肠杆菌中表达MBP-mEra蛋白。通过直链淀粉亲和层析纯化目标蛋白。用蛋白质免疫印迹法鉴定兔抗人Era抗体对这些蛋白的特异性。
表达的MBP-mEraW和MBP-mEraS蛋白分别约占细菌总蛋白的17%和19%。经直链淀粉亲和层析后，融合蛋白的纯度分别为67%和61%。兔抗人Era抗体对这两种剪接的小鼠Era蛋白具有高度特异性。
利用基因重组技术可在大肠杆菌中表达两种融合的mera基因。兔抗人Era抗体对两种剪接的小鼠ERA蛋白具有高度特异性，表明该抗体可用于研究这两种剪接的小鼠Era的功能。
