中文摘要：

目的：研究参与铁硫蛋白形成的关键分子BolA同系物3（BolA3）在细胞中的功能，通过重组表达BolA3蛋白，进而免疫制备并鉴定针对该分子的抗体。方法：从人肺癌细胞A549细胞中提取RNA，RT—PCR扩增出BolA3基因，构建BolA3基因重组原核表达载体pQE30-BolA3，使用IPTG诱导蛋白表达，Western blotting法鉴定表达中带His标签的重组BolA3蛋白，使用镍（Ni）柱纯化该蛋白。程序免疫大耳白兔，酶联免疫吸附试验（ELISA）方法测定兔血清中抗体滴度。Western blotting法检测制备的抗体与真核表达的BolA3蛋白结合的特异性，并利用已制备的BolA3蛋白抗体检测多种细胞中BolA3内源性表达。结果：克隆的BolA3基因序列与理论序列相一致，原核表达并纯化了BolA3重组蛋白，程序免疫大耳白兔后，获得了高滴度、高特异性的抗体血清。使用所得纯化的BolA3蛋白抗体检测到多种细胞内源性表达BolA3。结论：成功制备针对BolA3的多克隆抗体，为进一步深入研究BolA3蛋白的生物学功能奠定了基础。

英文摘要：

Objective：To prepare and identify the recombinant polyclone antibody against BolA homolog 3 （BolA3）, a key factor of iron-sulfur proleins formalion, and to study its function in cells. Methods： BolA3 was cloned from the cDNA which was amplified from the human hmg cancer A549 cells by RT-PCR, then subcloned into the prokaryotic expression vector pQE30. The expression of His-tagged BolA8 was induced by IPTG and purified by His Trap affinity column. The recombinant protein was used to immunize rabbit. The serum antibody titerin rabbits was detected by enzyme-linked immunosorbent assay （ELISA）. The specificity of antibody combined with BolA3 protein in several human cell lines were identified by western blotting. Results： The recombinant prokaryotic expression vector pQE30-BolA3 was successfully constructed. The high titer and high specificity of antibodics were obtained in rabbit serum. Western blotting showed that the purified BolA3 protein was expressed in several human cell lines. Conclusion. The polvclone anlibody against BolA3 was success- fully prepared,which laid the foundation for the further research of BolA3＇s biological function.