期刊论文

Expression of porcine epidemic diarrhea virus CH-HeB14 strain S1 protein and preparation of its polyclonal antibody

中文

西北农林科技大学学报(自然科学版)

1671-9387

2018

46

4

1-12

10.13207/j.cnki.jnwafu.2018.04.001

国家自然科学基金青年基金项目（31101837）； 衡阳师范学院***专项（16D20）； 湖南省教育厅优秀青年项目（17B041）；

本校为第一机构

【目的】制备猪流行性腹泻病毒(PEDV)CH-HeB14毒株S1蛋白的多克隆抗体,为PEDV新流行毒株诊断试剂盒和疫苗研制提供基础材料。【方法】利用RT-PCR扩增PEDV CH-HeB14毒株S1基因,对序列特征进行生物信息学分析,用PCR将S1基因截成不同片段(S1A、S1B、S1C、S1C-1和S1C-2),分别克隆至原核表达载体pET28a,转化BL21(DE3)构建重组表达菌株;用IPTG诱导重组蛋白表达,并进行SDS-PAGE和Western blotting鉴定。将表达蛋白与佐剂乳化制备免疫原免疫兔多克隆抗体,Western blotting和免疫过氧化物酶单层细胞染色法(IPMA)检测抗体的免疫活性。【结果】 PEDV S1、S1A、S1B、S1C、S1C-1和S1C-2片段长度依次为2 079,6...

【Objective】 This study prepared polyclonal antibody of S1 protein of porcine epidemic diarrhea virus (PEDV) CH-HeB14 strain to provide basis for the development of diagnostic kit and vaccine.【Method】 The S1 gene of PEDV CH-HeB14 strain was amplified by reverse transcription-polymerase chain reaction (RT-PCR),analyzed by bioinformatics software,truncated into different segments (S1A,S1B,S1C,S1C-1 and S1C-2) by PCR,sub-cloned into prokaryotic expression vector pET28a and transformed into BL21(DE3) to obtain the expression recombinants.IPTG wa...