免疫调控相关基因C12orf28在免疫组织中的分布特征  

Expression features of a novel immune-related gene C12orf28 in immunologic tissues

作　　者：任慧[1,2] 李红敏[1,2] 乔雍[1,2] 刘燃[1,2] 郝晓花[1,2] 魏红山[1,2] 

机构地区：[1]首都医科大学附属北京地坛医院传染病研究所 [2]新发突发传染病研究北京市重点实验室,北京100015

出　　处：《中华实验和临床感染病杂志（电子版）》2013年第6期1-4,共4页Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition)

基　　金：国家自然科学基金(No.81071411;No.81271901)

摘　　要：目的制备抗人C12orf28多克隆抗体,明确C12orf28编码蛋白在免疫组织中的分布特征。方法选取免疫原性高、特异性强的羧基端257个氨基酸作为目的片段,以人外周血单个核细胞(PBMC)cDNA为模板,采用PCR获得目的基因C12orf28C257;构建其原核表达载体pET32a(+)-C12orf28C257,转化入大肠埃希菌BL21(DE3),异丙基β-D-硫代吡喃半乳糖苷(IPTG)诱导表达重组蛋白;将重组蛋白免疫大耳白大白兔制备兔源性多克隆抗体,采用ELISA和Western blot分析检测多克隆抗体效价和特异性,采用Western blot明确C12orf28的组织分布特征。结果 PCR扩增得到C12orf28C257目的基因片段,诱导表达重组蛋白并制备了其多克隆抗体。ELISA分析结果显示,多克隆抗体效价>1︰1.28×106,Western blot分析鉴定得出多克隆抗体具有较高的特异性。结论未知功能基因C12orf28在胎儿肠、淋巴、胸腺和心肌等组织中均有不同程度的表达。Objective To prepare the polyclonal antibody of C12orf28 and observe the expression features of C12orf28 in immunologic tissues. Methods The peptide sequence with high immunogenicity and specificity that contains 257 amino acids of the carboxy-terminal was selected and amplified by reverse transcription PCR. The DNA fragment C12orf28C257 was inserted into pET32a（＋） plasmid and transformed into Escherichia coli BL21. The protein expression was induced with IPTG in vitro. Anti-C12orf28 was prepared by immunizing the big ears rabbit and was analyzed by Western blot and ELISA. The distribution of C12orf28 in tissues was detected by Western blot. Results The DNA fragment C12orf28C257 was cloned and its recombinant protein was successfully induced. The antibody titer was more than 1︰1.28 × 106 by ELISA and the high specificity of the polyclonal antibody was detected by Western blot. Conclusion The new gene C12orf28 is expressed in the fetal tissues such as intestine, lymph node, thymus and myocardiun, etc.

关 键 词：C12orf28 基因表达 重组蛋白 多克隆抗体 肝炎 乙型 

分 类 号：R392.1[医药卫生—免疫学]