中国兽医科学2018，Vol.48Issue(2)：155-160,6.DOI:10.16656/j.issn.1673-4696.2018.0024
禽源CCL19蛋白的表达纯化和抗体的制备
Expression of avian origin chemokine CCL19 and preparation of its polyclonal antibodies
摘要
Abstract
To study the effects of CCL19 on the development of avian disease,CCL19 was cloned from the bursae of IBDV-inoculated chickens by RT-PCR.Sequencing results showed that the cDNA of CCL19 contained a 300 bpopen reading frame(ORF),which encoded 100amino acids and shared 100％nucleotide acid and amino acid similarity with avian CCL19 deposited in GenBank.The cDNAof CCL19 was then subcloned into pET30a vector,transformed into E.coli BL21(DE3)and induced with IPTG.SDS-PAGE indicated that the molecular weight of the expressed fusion protein was approximately 13 ku,which was the same as the expected size.High purity recombinant protein was obtained by Ni-sepharose purification.Western-blot showed that the purified fusion protein could be recognized by the positive sera of mice against His-tag protein.The purified target protein was used to immunize New Zealand white rabbits and the obtained polyclonal antibodies were identified with CCL19 protein by Western-blot.In all,the preparations of avian origin chemokine CCL19 protein and its polyclonal antibodies have laid the foundation for studies of the development of avian disease.关键词
趋化因子/CCL19/鸡/表达/多克隆抗体Key words
chemokines/CCL19 gene/gallus/expression/polyclonal antibody分类
农业科技引用本文复制引用
王秋霞,李鹏祥,李茵,刘兴友,余燕,张艳红,姜金庆,马金友,欧长波..禽源CCL19蛋白的表达纯化和抗体的制备[J].中国兽医科学,2018,48(2):155-160,6.基金项目
国家自然科学基金项目(31402169) （31402169）
畜禽疫病防控河南省高校科技创新团队支持计划资助项目(18IRTSTHN019) （18IRTSTHN019）
河南科技学院2017年大学生创新创业训练项目(2017CX037) （2017CX037）
