南方医科大学学报2017，Vol.37Issue(3)：312-316,5.DOI:10.3969/j.issn.1673-4254.2017.03.06
奇异变形杆菌聚磷酸盐激酶多克隆抗体的制备和鉴定
Expression of Proteus mirabilis polyphosphate kinase and preparation of its polyclonal antibodies
摘要
Abstract
Objective To express and purify polyphosphate kinase (PPK) from Proteus mirabilis and prepare the polyclonal antibody against PPK.Methods The antigenicity and hydrophobicity of PPK were analyzed using software.The N-terminal conservative sequence containing 309 amino acids was selected as the target peptide,and its corresponding gene sequence with modification based on prokaryotic cells-preferred codon was synthesized and inserted into plasmid pET28b(+ ).The constructed recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and induced with IPTG.The expressed fusion protein was purified using Ni-affinity chromatography.The purified protein was injected along with adjuvant in rabbits to prepare the polyclonal antibodies against PPK.Results and Conclusion PPK fusion protein expressed by E.coli was purified successfully using Ni-affinity chromatography.ELISA result demonstrated that the harvested rabbit anti-sera against PPK had a high titer of 1:512 000,and Western blotting showed a good specificity of the antibody,which can be used further study of the role of PPK in the pathogenesis of Proteus mirabilis infection.关键词
奇异变形杆菌/聚磷酸盐激酶/多克隆抗体Key words
Proteus mirabilis/ polyphosphate kinase/ polyclonal antibody2016-10-05引用本文复制引用
彭亮,区静怡,潘嘉韵,邓聪,陈景红,曹虹..奇异变形杆菌聚磷酸盐激酶多克隆抗体的制备和鉴定[J].南方医科大学学报,2017,37(3):312-316,5.基金项目
国家自然科学基金(81670637) （81670637）
广州市属高校科技项目(1201430488) （1201430488）
广州医科大学青年项目(2014A06)Supported by National Natural Science Foundation of China (81670637). （2014A06）
