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A Human-Specific De Novo Protein-Coding Gene Associated with Human Brain Functions

Figure 5

Western-blot assays to determine the protein expression levels of FLJ33706.

(A) A specific band with molecular mass of about 22 kDa in SDS-PAGE, which was consistent with the predicted molecular weight of FLJ33706 putative proteins, was detected in the Western-blot assay. The band could not be detected in pre-immune control and peptide competition control. Pre: pre-immune reaction assay; Ab: FLJ33706 antibody assay; Pep: peptide competition assay. (B) E. coli samples before and after the transformation of FLJ33706 recombination plasmids were assayed by Western blot using (a) His-tag specific antibody and (b) anti-FLJ33706. FLJ33706 expression in human cortex was shown in (c) as the control. Before: E. coli samples before the plasmid transformation; After: E. coli samples after the plasmid transformation. (C) The specific band can be detected in all human brain regions, but not in mouse brain regions used as controls. H-CTX, M-CTX: human/mouse cortex; H-MID, M-MID: human/mouse midbrain; H-CER, M-CER: human/mouse cerebellum; (D) FLJ33706 expression can be detected in different human individuals. 384, 994, 275, 995, 705, 710, 1277: individual IDs. 22KD: theoretical molecular weight of FLJ33706 protein; 42KD: molecular weight of beta-actin protein as endogenous control. In A and C, antibodies for FLJ33706 and endogenous control were mixed in Western assays.

Figure 5