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为从细胞水平上研究独角雪冰鱼Chionodraco hamatus和伯氏肩孔南极鱼Trematomus bernacchii的低温适应能力,采用RT-PCR方法,获得独角雪冰鱼和伯氏肩孔南极鱼的瞬时受体电位阳离子通道蛋白(transient receptor potential channels V1,TRPV1)基因序列,其开放阅读框全长均为2310 bp;生物信息学分析表明,两种南极鱼TRPV1基因均编码769氨基酸残基,预测其蛋白相对分子质量分别为88 210和88 160,理论等电点分别为7.63和8.26;两种南极鱼TRPV1蛋白均无信号肽;亚细胞定位显示,TRPV1蛋白主要在细胞膜和内质网中发挥生物学功能;理化性质分析表明,两种南极鱼的TRPV1为不溶性跨膜蛋白,分别有34、35个磷酸化位点和6、5个糖基化位点,二者均含有6个跨膜结构域、5个锚蛋白结构域,二级结构以随机卷曲为主;同源性分析显示,独角雪冰鱼TRPV1氨基酸序列与伯氏肩孔南极鱼的一致性最高,为93.4%,与其他物种的一致性为44.3%~69.8%;低温适应试验结果显示,两种南极鱼的TRPV1基因不能被温度所激活。研究表明,独角雪冰鱼和伯氏肩孔南极鱼TRPV1基因可能与低温适应无关。
In order to study the hypothermic adaptability of Chionodraco hamatus and Trematomus bernacchii from the cellular level, we obtained the transient receptor of Antarctic iceworms The full-length open reading frame of TRPV1 gene was 2310 bp in length. The bioinformatics analysis showed that the TRPV1 genes of two Antarctic fish all encoded 769 amino acid residues, which predicted the protein relative The molecular weights were 88 210 and 88 160, respectively. The theoretical isoelectric points were 7.63 and 8.26, respectively. No signal peptide was found in the two Antarctic fish TRPV1 proteins. Subcellular localization showed that TRPV1 protein mainly played a biological role in the membrane and endoplasmic reticulum ; Physicochemical analysis showed that TRPV1 of two Antarctic fish were insoluble transmembrane proteins with 34,35 phosphorylation sites and 6,5 glycosylation sites, respectively, and both contained six transmembrane domains, Five ankyrin domains with secondary structure mainly randomized. Homology analysis showed that the TRPV1 amino acid sequence of Unicornis fulgiformis had the highest identity (93.4%) with that of the shoulder shoulder Antarctic fish, Consistent The results showed that the TRPV1 gene of two Antarctic fish could not be activated by temperature. Studies have shown that Antarctic fish Antarctic icefish and Berberis TRPV1 gene may be cold adaptation has nothing to do.