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以中华鲟(Acipenser sinensis)脑垂体总RNA为模板,采用RT-PCR和RACE方法,获得中华鲟神经内分泌多肽(7B2)基因的3个重叠片段,测序后拼接得到986 bp全长基因序列,其中包括5’端非翻译区(5′-UTR)14 bp、3′端非翻译区(3-′UTR)261 bp和开放阅读框711 bp。翻译编码236个氨基酸。其中前43个氨基酸为7B2的信号肽。经BLAST比对发现中华鲟7B2蛋白的同源性与斑马鱼(Danio rerio)的相似性最高为82%。系统发育分析表明,中华鲟与斑马鱼亲缘关系最近。半定量RT-PCR分析表明:在脑中7B2 mRNA表达量最高,心脏、性腺、胰等组织中表达次之,肠、肾、皮肤等组织中少量表达,肝和鳃几乎不表达。
Three overlapping fragments of 7B2 gene of Chinese sturgeon were obtained by RT-PCR and RACE using total RNA of the pituitary gland of Acipenser sinensis as a template. After sequencing, the full-length cDNA sequence of 986 bp was obtained, Including 14 bp 5 ’untranslated region (5’-UTR), 261 bp 3’ untranslated region (3 -UTR) and 711 bp open reading frame. Translation encodes 236 amino acids. The first 43 amino acids were 7B2 signal peptide. The BLAST alignment showed that the similarity of 7B2 protein of Chinese sturgeon with Danio rerio was 82%. Phylogenetic analysis showed that the Chinese sturgeon has the closest genetic relationship with zebrafish. Semi-quantitative RT-PCR analysis showed that expression of 7B2 mRNA was the highest in brain, followed by expression in heart, gonad, and pancreas, followed by small expression in gut, kidney and skin, with little expression in liver and gill.