文献类型：期刊文献

中文题名：禽源趋化因子CCL4基因的表达及多克隆抗体的制备

英文题名：Expression of avian origin chemokine CCL4 and preparation of its polyclonal antibodies

作者：王秋霞[1];王芳[1];楚富茗[1];魏小兵[1];胡会龙[1];刘兴友[1,2];余燕[1];张艳红[1];马金友[1];姜金庆[1];欧长波[1]

第一作者：王秋霞

机构：[1]河南科技学院动物科技学院,河南新乡453003;[2]新乡学院,河南新乡453003

第一机构：河南科技学院动物科技学院,河南新乡453003

年份：2020

卷号：50

期号：1

起止页码：93-98

中文期刊名：中国兽医科学

外文期刊名：Chinese Veterinary Science

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD_E2019_2020】；

基金：国家自然科学基金项目(31402169);河南省科技攻关项目(182102110301,192102110189);河南省高校科技创新团队支持计划资助项目(18IRTSTHN019)

语种：中文

中文关键词：趋化因子;CCL14;禽源;表达;多克隆抗体

外文关键词：Chemokines;CCL4 gene;avian origin;expression;polyclonal antibodies

摘要：在禽源疾病发生发展过程中,趋化因子CCL4的作用尚不清楚。对克隆的禽源CCL4基因序列分析显示,其ORF包含270个碱基,编码90个氨基酸,与GenBank中登录的禽源CCL4序列完全一致;利用原核表达载体pET30a对CCL4的成熟肽部分进行表达,IPTG诱导后,进行SDS-PAGE电泳,结果与预期一致,在约11 ku处出现明显蛋白条带。使用经镍柱纯化获得的目的蛋白免疫家兔,制备多克隆抗体。经Western-blot检测,获得的抗体能与CCL4蛋白发生特异性反应。禽源CCL4蛋白的首次克隆、表达和纯化,以及抗CCL4蛋白多克隆抗体的制备,为进一步研究趋化因子CCL4在禽源疾病发生发展过程中的作用奠定了基础。
The role of chemokine CCL4 in the development of avian diseases remains unclear.The CCL4 gene was cloned and the sequencing results showed that its open reading frame(ORF)contained 270 bases and encoded 90 amino acids.The sequence shared 100%nucleotide acid and amino acid similarity with avian CCL4 deposited in Gen Bank.The CCL4 cDNA fragment without signal peptide was expressed by the prokaryotic expression vector pET30 a.After IPTG induction,SDS-PAGE result indicated the band was consistent with the expectation at about 11 ku.Polyclonal antibodies were prepared by immunizing rabbits with purity protein by Ni-sepharose purification.Western-blot analysis showed that the antibody could react with CCL4 protein.In all,the expression and purification of avian origin chemokine CCL4 protein,and the preparation of anti-CCL4 protein polyclonal antibodies,have laid the foundation for further research on the role of chemokine CCL4 in development of avian disease.