一种与HBx蛋白相互作用未知蛋白抗体的制备及其应用

一种与HBx蛋白相互作用未知蛋白抗体的制备及其应用 Preparation and Application of Antibody for Unknown Protein Interacting with HBx Protein 张建林 ZHANG Jian-lin 1 2 first-author 思维 SI Wei 1 2 赵伟光 ZHAO Wei-guang 1 2 黄静 HUANG Jing 1 2 朱应 ZHU Ying 1 2 李雁 LI Yan 1 2 章晓联 ZHANG Xiao-lian 1 2 吴建国 WU Jian-guo 1 2 武汉大学生命科学学院/病毒学国家重点实验室 武汉大学生命科学学院/病毒学国家重点实验室 College of Life Sciences,Wuhan University/State Key Laboratory of Virology College of Life Sciences,Wuhan University/State Key Laboratory of Virology 武汉大学生命科学学院/病毒学国家重点实验室,湖北,武汉,430072 武汉大学生命科学学院/病毒学国家重点实验室,湖北,武汉,430072 本研究构建UK蛋白的原核表达载体pET28-UK,转化BL21宿主菌,经过IPTG诱导、表达、纯化,获取UK蛋白,制备抗原,免疫杭州大耳白兔.经过3次免疫后心脏采血,吸取血清作为抗体.经过ELISA和免疫荧光试验验证,该抗体可以识别机体UK蛋白.本研究为深入阐明HBx与UK蛋白的相互作用的功能性试验,以及完全阐明UK蛋白在细胞内的功能奠定了基础. The DNA fragment including 3′-terminate of UK gene was amplified from plasmid pOT-UK by PCR and cloned into plasmid pET28;the new plasmid was named pET28-UK.The plasmid pET28-UK was transformed into E coli.BL21.Protein UK was expressed by inducing transformed BL21 with IPTG and purified by ringing using PBS.The protein UK mixed with completed Ford’s adjuvant was first injected rabbits,and then injected with UK protein with uncompleted Ford’s adjuvant.10 days later after last immunity with UK protein,the serum of rabbits was extracted from heart.The antibody can identify the protein UK in the cells by immunofluorescence. HBx蛋白 UK蛋白 抗体 应用 HBx protein UK protein antibody application R392.1 国家自然科学基金资助项目（30570070） 2005-06-01 2021-04-01 6