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요약정보
猪肺炎支原体脂蛋白LppT的多克隆抗体制备及鉴定
Resource Type
Academic Journal
Authors
刘威
;
袁芳艳
;
周丹娜
;
刘泽文
;
杨克礼
;
段正赢
;
郭锐
;
蔡行
;
肖少波
;
田永祥
Source
江苏农业科学 / Jiangsu Agricultural Sciences
. 45(19):220-223
Subject
猪肺炎支原体
脂蛋白
LppT
多克隆抗体
Language
Chinese
ISSN
1002-1302
Online Access
Find it @ DONGA
초록
试验构建猪肺炎支原体重组表达质粒pET30a-LppT,通过快速定点突变试剂盒将LppT基因中513、945、1 860、2 547 bp处的A突变为G,使改造后的LppT基因能在大肠杆菌系统中正确表达,IPTG诱导重组蛋白的表达并进行了蛋白纯化,纯化的目的蛋白与佐剂混合、乳化制备免疫原,免疫新西兰大白兔制备LppT蛋白多克隆抗体.SDS-PAGE分析结果显示,IPTG诱导表达后获得1条特异性蛋白条带,分子质量约为110 ku,与预期蛋白大小一致.可溶性分析结果表明目的蛋白以可溶形式表达于上清中,纯化的蛋白经SDS-PAGE分离后,用制备的LppT多克隆抗体进行Western Blot检测,结果表明制备的LppT多克隆抗体具有较强的特异性.
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