The Bicaudal-C (Bic-C) gene was first discovered in Drosophila melanogaster, it’s normal function is required for correct targeting of the migrating anterior follicle cells and for anterior-posterior patterning of the Drosophila oocyte. Mouse Bicc1 gene is a homologe of Drosophila Bicaudal-C gene, whose mutation causes a phenotype sembles with human polycystic kidney disease, but the details of Bicc1 involved in cystogenesis are still elusive. Here we cloned the full-length cDNA sequence of Bicc1 gene by nest PCR and digest- ligate methods. Based on the bioinformatics analysis results of mouse Bicc1, the DNA fragment encoding N-terminal 139 amino acids(61E-199A) and C-terminal 148 amino acids(711Q-858D) of Bicc-1 was obtained by PCR and was cloned into a prokaryotic GST-fusion protein expression vector. Fusion protein were expressed after IPTG induction, and then purified from the total proteins of E. coli Rosetta cells transformed by the recombinant plasmids pGex-HisC3-GST- Bicc1-N/C. We prepared two polyclone antibodies against mouse Bicc1 protein with high specificity by immunizing rabbit with these two purified fusion proteins. The specificity of the affinity-purified antibodies was iedntified by Western blot. We stained cultured cells and mouse kidney paraffin sections with this antibody and found that Bicc1 is mainly localized at cytoplasm in cultured mouse kidney cells. And Bicc1fristly just located in heart and neural tube in early mouse embryo, then found in many different tissues and keep stable expression level. Bicc1 mDNA expess in different organs and mainly in kidney. Get two RNAi sequences that specifically knock-down Bicc1expression level in cultured cells by fluorescence and Western-blot, provide the tools for the research in low-expression stable cell lines.
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