摘要
根据GenBank中报道的鸡GABA(A)α1一级结构信息,用TMHMM 2.0和DNAstar软件分析GABA(A)α1蛋白的跨膜结构域、二级结构、疏水性、亲水性以及抗原性等,综合考虑抗体设计的其他因素,设计出一段16个氨基酸的多肽。将合成后的多肽与牛血清白蛋白(BSA)偶联,用与BSA偶联的GABA(A)α1多肽免疫新西兰兔,3个月后获取血清,亲和纯化GABA(A)α1多肽抗体。通过ELISA法检测抗体效价,Western blot和免疫组化法检测抗体特异性。通过该方法得到了高效价与高特异性的鸡GABA(A)α1多肽抗体,ELISA法测定其效价可达到1∶128 000,而且该抗体可用于免疫组化,说明所制备的鸡GABA(A)α1抗体具有高特异、高效价等特点,将为鸡GABA(A)α1基因的功能研究提供有用的研究材料。
According to the primary structure information in GenBank, the prediction of the possible conformational structure, hydrophobic-ity and antigenicity of chicken GABA（A） α1 with TMHMM 2. 0 and DNAstar software, and the principal for antibody production, a partial peptide with 16-amino acid residues of GABA（A） α1 was synthesized. The synthesized peptide was then used to immunize New Zealand rabbit after coupled with BSA. After 3 months, the serum was collected and the antibody to GABA（A） ctl peptide was purified. The titer and specificity of anti-GABA（A） ctl peptide antibody were determined by ELISA, Western blot and immunohistochemical analyses. The chicken GABA（A） α1 peptide antibody with high specificity was obtained and the titer could achieve 1 ： 128 000 by ELISA. And the anti-body could be used in the immunohistochemical analysis. This study will provide useful material for function research of GABA（A） α1 gene.
出处
《畜牧与兽医》
北大核心
2013年第11期1-5,共5页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31172242)
