CXXC5多克隆抗体的制备及表达研究

王西君; 廖鹏; 王跃群; 邓云; 莫小阳; 袁婺洲; 万永奇; 吴秀山; 李永青

doi:doi:10.3969/j.issn.1007-7146.2011.03.011

激光生物学报, 2011, 20 (3): 334, 网络出版: 2015-10-08

CXXC5多克隆抗体的制备及表达研究

A Preparation of Polyclonal Antibody and Expression of Human CXXC5 Gene

论文大纲

王西君廖鹏王跃群邓云莫小阳袁婺洲万永奇吴秀山李永青 ^*

作者单位

湖南师范大学蛋白质化学及鱼类发育生物学教育部重点实验室, 心脏发育研究中心, 湖南长沙 410081

融合蛋白多克隆抗体 CXXC5 CXXC5 fusion protein polyclonal antibody

摘要

CXXC5基因是从人类胚胎心脏的cDNA文库中克隆出来的一个人类锌指基因, 包含zf-CXXC5结构域, 该基因编码322个氨基酸, 在物种进化上高度保守。为了进一步研究该基因的功能, 需要获得CXXC5蛋白并制备其抗体.通过PCR扩增方法扩增得到了CXXC5部分编码区序列, 然后将其连接到PGEX-4T-1 上, 转化到大肠杆菌BL21 (DE3)中, 再通过自身诱导法诱导表达重组质粒的融合蛋白. 通过割胶回收纯化融合蛋白。免疫新西兰大白兔制备多克隆抗体, Western blot 检测抗体活性。结果表明, 实验获得了高质量的多克隆抗体。

Abstract

CXXC5 was cloned from a human heart cDNA library and identified as human zf-CXXC-related zinc finger gene encoding a 322 amino acid protein. CXXC5 is an evolutionarily conserved protein. Obtianing the CXXC5 protein and it’s antibody is essential for the further studies. A cDNA fragment of partial coding sequence of CXXC5 was amplified by PCR, then the PCR product was cloned into the expression vector pGEX-4T-1 and transformed into Escherichia coli BL21 (DE3). The fusion protein of the recombinant expression plasmid was expressed by a way of auto-induction. The fusion protein was separated with SDS-PAGE and recovered by gel extraction. New Zealand big white rabbits were immunized with the separated protein and the antibody titer and specification was identified by Western blot. The result showed that the high quality multi-clonal antibody was obtained.

参考文献

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王西君, 廖鹏, 王跃群, 邓云, 莫小阳, 袁婺洲, 万永奇, 吴秀山, 李永青. CXXC5多克隆抗体的制备及表达研究[J]. 激光生物学报, 2011, 20(3): 334. WANG Xi-jun, LIAO Peng, WANG Yue-qun, DENG Yun, MO Xiao-yang, YUAN Wu-zhou, WAN Yong-qi, WU Xiu-shan, LI Yong-qing. A Preparation of Polyclonal Antibody and Expression of Human CXXC5 Gene[J]. Acta Laser Biology Sinica, 2011, 20(3): 334.

CXXC5多克隆抗体的制备及表达研究

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