动物医学进展
動物醫學進展
동물의학진전
PROGRESS IN VETERINARY MEDICINE
2014年
6期
11-15
,共5页
廖娟红%贾广乐%乔海燕%林祥梅
廖娟紅%賈廣樂%喬海燕%林祥梅
료연홍%가엄악%교해연%림상매
贝氏柯克斯体%精氨酸受体%原核表达%免疫原性
貝氏柯剋斯體%精氨痠受體%原覈錶達%免疫原性
패씨가극사체%정안산수체%원핵표체%면역원성
Coxiella burnetii%arginine repressor(argR)%prokaryotic expression%immunogenicity
构建贝氏柯克斯体精氨酸受体（argR）的重组表达载体并分析该重组蛋白的免疫原性。应用PCR 方法，以贝氏柯克斯体九里株为模板，扩增出含734 bp 的精氨酸受体目的基因片段，并将其克隆至原核表达载体 pGEX-6p-1，得到重组表达质粒 pGEX-6p/argR，经 IPTG 诱导表达后产生 ArgR 重组蛋白，SDS-PAGE 分析该蛋白大小约为25 ku，Western blot 结果显示，表达的重组蛋白有着良好的免疫原性。用 argR免疫小鼠，能诱发特异性抗体的产生，说明该原核表达蛋白具有良好的免疫原性。
構建貝氏柯剋斯體精氨痠受體（argR）的重組錶達載體併分析該重組蛋白的免疫原性。應用PCR 方法，以貝氏柯剋斯體九裏株為模闆，擴增齣含734 bp 的精氨痠受體目的基因片段，併將其剋隆至原覈錶達載體 pGEX-6p-1，得到重組錶達質粒 pGEX-6p/argR，經 IPTG 誘導錶達後產生 ArgR 重組蛋白，SDS-PAGE 分析該蛋白大小約為25 ku，Western blot 結果顯示，錶達的重組蛋白有著良好的免疫原性。用 argR免疫小鼠，能誘髮特異性抗體的產生，說明該原覈錶達蛋白具有良好的免疫原性。
구건패씨가극사체정안산수체（argR）적중조표체재체병분석해중조단백적면역원성。응용PCR 방법，이패씨가극사체구리주위모판，확증출함734 bp 적정안산수체목적기인편단，병장기극륭지원핵표체재체 pGEX-6p-1，득도중조표체질립 pGEX-6p/argR，경 IPTG 유도표체후산생 ArgR 중조단백，SDS-PAGE 분석해단백대소약위25 ku，Western blot 결과현시，표체적중조단백유착량호적면역원성。용 argR면역소서，능유발특이성항체적산생，설명해원핵표체단백구유량호적면역원성。
This study is aimed to construct and identify the prokaryotic expression plasmids of arginine re-pressor gene(argR)of Coxiella burnetii and analyze its immunogenicity.The argR from Nine Mile strain was amplified by PCR,argR fragment is 734 bp.The argR gene was cloned into the prokaryotic expression vector pGEX-6p-1 after identification by PCR,enzyme digestion and sequencing.The recombinant plasmid pGEX-6p/argR was constructed,and then the plasmid was transformed into BL21(DE3)competent cells and induced by IPTG.The result showed that pGEX-6p/argR recombinant protein is 25 ku.SDS-PAGE and Western blot indicated that the recombinant protein could express argR protein with significant antigenci-ty.The protein argR induced specific antibodies in immunized mice and had good immunoreactivity.
