中文摘要：

制备鸡（Gallus gallus）肝脏胆汁酸结合蛋白（L-BABP）的多克隆抗血清,并分析L-BABP的组织表达特性。利用RT-PCR扩增L-BABP基因的CDS区,构建鸡L-BABP基因的GST融合蛋白表达质粒pGEX-4T/L-BABP。将重组表达质粒转化到大肠杆菌（Escherichia coli）BL21中,经IPTG诱导后产生GST/L-BABP融合蛋白,利用GlutathioneSepharose4B亲和层析纯化目的蛋白,将纯化的GST/L-BABP融合蛋白免疫家兔制备多克隆抗血清。诱导得到了1个38kD（12kDL-BABP＋26kDGST）的融合蛋白,经间接ELISA方法测定制备的抗血清效价为1：100000。利用此抗血清分析鸡L-BABP的组织表达特性,结果表明该基因仅在肝脏组织中特异性表达,在肾、肺、腿肌、腺胃、胸肌、心脏、脂肪、脾、回肠、肌胃、空肠和十二指肠等12种组织中没有检测到表达信号。本研究制备的高效价、高特异性的鸡L-BABP抗血清,为从蛋白水平上深入研究L-BABP的生物学功能提供了良好的基础。

英文摘要：

The aim of this study is to prepare the antiserums against chicken liver bile acid binding protein （L-BABP） and analyze expression characteristics of L-BABP.Specific primers were designed to amplify the coding region of chicken L-BABP by RT-PCR.And the L-BABP gene was then inserted into pGEX-4T-1 vector and expressed in Escherichia coli BL21 （DE3） with IPTG induction.Then the fusion protein was purified by Glutathione Sepharose 4B affinity chromatography and the antiserums against L-BABP was produced by immunizing rabbits.The results showed that a 38 kD （12 kD L-BABP＋26 kD GST） fusion protein was induced.And the titer of the antiserum against GST/L-BABP was 1：100 000 detected by indirect ELISA.Analysis of tissue expression showed that L-BABP specificly expressed in liver,and there were no detectable signal in kidney,lung,crureus,glandular stomach,pectorales,heart,fat,spleen,ileum,muscle stomach,jejunum and duodenum,respectively.In the current study,preparation of high titer and high specificity of the chicken L-BABP antiserum provides a good foundation for studying the biological function of L-BABP in the protein level.