鸡L-FABP抗血清制备及组织表达特性分析  

Preparation of Antiserums against Chicken Liver-type Fatty Acid Binding Protein (L-FABP) and Tissue Expression Analyses of L-FABP

作　　者：石慧[1] 王启贵[1] 王宇祥[1] 王宁[1] 李辉[1] 

机构地区：[1]东北农业大学动物科学技术学院,哈尔滨150030

出　　处：《畜牧兽医学报》2008年第11期1466-1469,共4页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金重点项目(30430510);黑龙江省普通高等学校青年学术骨干支持计划项目(1152G005);国家高技术研究发展计划(863)项目(2006AA10A120)

摘　　要：为制备鸡肝脏型脂肪酸结合蛋白(L-FABP)的多克隆抗血清,并分析L-FABP的组织表达特性,利用RT-PCR扩增L-FABP基因,构建鸡L-FABP基因的GST融合蛋白表达质粒pGEX-4T/L-FABP。将重组表达质粒转化大肠杆菌BL21,IPTG诱导产生GST/L-FABP融合蛋白,用亲和层析纯化目的蛋白,将纯化的GST/L-FABP融合蛋白免疫家兔制备抗血清,并利用此抗血清分析鸡L-FABP基因的组织表达特性。诱导得到了1个40 ku(14ku L-FABP+26 ku GST)的融合蛋白,获得效价较高、特异性强的鸡L-FABP的抗血清。鸡L-FABP的组织表达特性研究结果表明,该基因在肝脏和小肠组织中有较高表达,但在心脏、脂肪、肌肉、肌胃、脾、肺和肾中没有检测到表达信号。The aim of this study was to prepare the antiserums against chicken liver-type fatty acid binding protein （L-FABP） and analyze expression characteristics of L-FABP. Specific primers were designed to amplify the gene of chicken L-FABP by RT-PCR. The L-FABP gene was then inserted into pGEX4T-1 vector and expressed in Escherichia coli BL21 （DE3）. The protein was purified by Glutathione Sepharose 4B affinity chromatography and the antiserums against L- FABP was produced by immunizing rabbits. The tissue expression characteristics of L-FABP gene were determined by Western blot analysis. The results showed that a 40 ku （14 ku L-FABP ＋ 26 ku GST） fusion protein was induced, furthermore, high titer antiserums against L-FABP was obtained. The tissue expression analysis showed that L-FABP was highly expressed in liver and small intestine but no hybridization signal was detected in heart, fat, muscle, muscle stomach, spleen, lung and kidney.

关 键 词：原核表达 抗血清 鸡 L-FABP 

分 类 号：S852.5[农业科学—基础兽医学] Q786[农业科学—兽医学]