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| Antiserums Preparation and Tissue Expression Analyses of Chicken Liver Bile Acid Binding Protein(L-BABP) |
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Abstract 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.
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Received: 10 May 2010
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