福建农林大学学报（自然科学版）2016，Vol.45Issue(3)：290-295,6.DOI:10.13323/j.cnki.j.fafu(nat.sci.).2016.03.009
巨型艾美耳球虫IMP1基因的原核表达及多克隆抗体的制备
Prokaryotic expression and preparation of polyclonal antibody against IMP1 protein of Eimeria maxima
摘要
Abstract
Immune mapped protein-1 ( IMP1) , found in Eimeria maxima, demonstrates excellent immunogenic and immunoprote-cive property. To make full use of this property in coccidiosis prevention, manufacture of polyclonal antibody against EmIMP1 was started with obtaining EmIMP1 gene by RNA extraction from unsporulated oocysts of E.maxima XinJiang Strain and cDNA synthesis by reverse transcription. Then amplified fragment of EmIMP1 gene was inserted into prokaryotic expression vector to construct ex-pression plasmid pET-28a-EmIMP1, after that the recombinant vector was transformed into BL21(DE3) E.coli for protein expres-sion. Serum was obtained by subcutaneously injecting the purified protein with the same amount of adjuvant in 2 rabbits every 2 weeks for 4 times. Finally, Western-blot was applied to determine the specificity of the protein and enzyme-linked immuno sorbent assay ( ELISA) was used to test the titer of the polyclonal antibody. Results showed that EmIMP1 gene was 140 bp in length and the pressed protein was 70 ku. The protein existed in both supernatant and inclusion body. The polyclonal antibody demonstrated good specificity and high titer. To summarize, IMP1 produced in rabbit was promising to play a protective role in coccidiosis prevention.关键词
巨型艾美耳球虫/IMP1基因/原核表达/多克隆抗体Key words
Eimeria maxima/IMP1 gene/prokaryotic expression/polyclonal antibody分类
农业科技引用本文复制引用
林倩,黄驹辉,唐牧之,江和基,黄志坚,殷光文..巨型艾美耳球虫IMP1基因的原核表达及多克隆抗体的制备[J].福建农林大学学报（自然科学版）,2016,45(3):290-295,6.基金项目
国家自然科学基金---青年基金项目(31502058)；福建省科技厅引导性项目(2015N0017)；大学生创新创业训练计划项目(201510389187) （31502058）
