应用与环境生物学报
應用與環境生物學報
응용여배경생물학보
CHINESE JOURNAL OF APPLIED & ENVIRONMENTAL BIOLOGY
2009年
5期
634-637
,共4页
赵巧香%张秀玲%纪丽丽%金福厚%计红%杨焕民%李士泽
趙巧香%張秀玲%紀麗麗%金福厚%計紅%楊煥民%李士澤
조교향%장수령%기려려%금복후%계홍%양환민%리사택
鼠%冷诱导RNA结合蛋白%多克隆抗体%基因重组
鼠%冷誘導RNA結閤蛋白%多剋隆抗體%基因重組
서%랭유도RNA결합단백%다극륭항체%기인중조
mice%CIRP%polyclonal antibody%gene recombination
冷诱导RNA结合蛋白(CIRP)是第一种在哺乳动物细胞中被发现的冷休克蛋白,伴随着细胞冷应激过程过量表达.为进一步研究冷诱导RNA结合蛋白CIRP的生物学功能,构建重组BALB/C鼠CIRP的原核表达载体,重组BALB/C鼠CIRP蛋白经NI-NTA亲和树脂纯化后免疫獭兔制备抗CIRP多克隆抗体,以间接ELISA和Western blot检测抗体效价和抗原特异性.本实验成功构建了CIRP的原核表达载体,在E coli XL-1-blue得到了高效表达,利用原核表达包涵体蛋白免疫獭兔获得了高效价的CIRP多克隆抗体,该抗体能识别肝脏和睾丸中天然的CIRP蛋白.图5参14
冷誘導RNA結閤蛋白(CIRP)是第一種在哺乳動物細胞中被髮現的冷休剋蛋白,伴隨著細胞冷應激過程過量錶達.為進一步研究冷誘導RNA結閤蛋白CIRP的生物學功能,構建重組BALB/C鼠CIRP的原覈錶達載體,重組BALB/C鼠CIRP蛋白經NI-NTA親和樹脂純化後免疫獺兔製備抗CIRP多剋隆抗體,以間接ELISA和Western blot檢測抗體效價和抗原特異性.本實驗成功構建瞭CIRP的原覈錶達載體,在E coli XL-1-blue得到瞭高效錶達,利用原覈錶達包涵體蛋白免疫獺兔穫得瞭高效價的CIRP多剋隆抗體,該抗體能識彆肝髒和睪汍中天然的CIRP蛋白.圖5參14
랭유도RNA결합단백(CIRP)시제일충재포유동물세포중피발현적랭휴극단백,반수착세포랭응격과정과량표체.위진일보연구랭유도RNA결합단백CIRP적생물학공능,구건중조BALB/C서CIRP적원핵표체재체,중조BALB/C서CIRP단백경NI-NTA친화수지순화후면역달토제비항CIRP다극륭항체,이간접ELISA화Western blot검측항체효개화항원특이성.본실험성공구건료CIRP적원핵표체재체,재E coli XL-1-blue득도료고효표체,이용원핵표체포함체단백면역달토획득료고효개적CIRP다극륭항체,해항체능식별간장화고환중천연적CIRP단백.도5삼14
Cold inducible RNA-binding protein (CIRP) is the first cold shock protein found in mammalian cells and overexpressed upon a temperature downshift. To further study the biological function of CIRP, polyclonal antibody of CIRP was prepared. The prokaryotic expression vector of recombination BALB / C mice CIRP was constructed, and recombinant protein CIRP was purified with the NI-NTA resin. The purified protein was used to generate polyclonal antibody in rabbits. CIRP antibody titer and antigen specificity were detected by indirect ELISA and Western blot test, respectively. The results showed that the prokaryotic expression vector of recombination BALB / C mice CIRP was constructed successfully and was highly expressed in E. coli XL-1-blue. High antibody titer polycional antibody was obtained, It could recognize natural CIRP protein in liver and testis. So, it is concluded that the high antibody titer polyclonal antibody is prepared and it can be combined with natural CIRP protein in liver and testis, which is useful for further studying the biological function of CIRP. Fig 5, Ref 14