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作 者:杨辉[1] 艾闽[1] 李锋刚[1] 刘小林[1,2] 王立新[1,2]
机构地区:[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]陕西省农业分子生物学重点实验室,陕西杨凌712100
出 处:《家畜生态学报》2012年第6期11-15,共5页Journal of Domestic Animal Ecology
基 金:陕西省科技攻关(2012K01-18);西北农林科技大学基本科研业务费专项(QN2011062);西北农林科技大学后稷学者支撑计划(Z11021007)
摘 要:从大鲵皮肤cDNA文库中分离获得大鲵α1-血红蛋白基因全长cDNA序列,生物信息学分析表明,大鲵α-血红蛋白基因全长cDNA序列为594bp,开放阅读框共432bp,编码144个氨基酸,编码的大鲵α1-血红蛋白推测的理论分子量为16.05kDa,等电点为6.44。氨基酸组成中酸性氨基酸11.9%,中性氨基酸69.9%,碱性氨基酸18.2%。结构分析表明,该蛋白没有信号肽,但在100~122个氨基酸处以及98~122个氨基酸处分别有由里向外、由外向里的跨膜螺旋区,二级结构以α-螺旋为主。氨基酸序进行同源性列比对表明与人、猕猴的亲缘关系最近,和牛蛙的亲缘关系最低只有35%。荧光定量PCR结果表明,α1-血红蛋白基因在肌肉中表达量最高,在胃中最低。In this case, α1-Hb cDNA sequence was isolated and identified from skin cDNA library of Chinese giant salamander. The α1-Hb gene was 594 bp in the full length with the ORF 432 bp. Besides, 144 kinds of amino acids were encoded byal-Hb gene, and the molecular weight was 16 048. 3 u in general, isoelectric point was 6.44. In addition, the structure analysis of α1-Hb showed that the composition of amino acids were acidic amino acids 11.9 %, neutral amino acids 69.9 % and basic amino acid 18.2 %. Structural analysis indicated that there was no signal peptide was existed in this protein but there was an outward transmembrane helical region from 100th to 122nd aa and also an inward transmembrane helical region from 98th to 122nd aa, whose secondary structure was mainly a-helix. The three-level structure of α1-HB protein indicated the evolutional relationship was that Andrias davidianus was homologous to land animals, with Homo sapiens and Macaca mulatta homology of α1-HB was nearly 50%, and Rana catisbianna took the lowest homology (35%). Moreover, the expression of α1-Hb gene mRNA in 7 tissues for Andrias davidianus were analyzed by the data of qRT-PCR, it was more expressed in muscle than the other six issues.
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