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西藏野生甜荞碳酸酐酶基因FsCA1的克隆和生物信息学分析
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作者 侯维海 王建林 +1 位作者 旦巴 胡单 《江苏农业科学》 北大核心 2017年第10期20-23,共4页
依据已公布的荞麦转录组测序信息,从Contig文库中获得了1个碳酸酐酶(carbonic anhydrase,简称CA)转录本,通过逆转录PCR(RT-PCR)扩增得到CA基因全长序列。生物信息学分析表明,FsCA1基因全长1 233 bp,开放阅读框978 bp,编码325个氨基酸;... 依据已公布的荞麦转录组测序信息,从Contig文库中获得了1个碳酸酐酶(carbonic anhydrase,简称CA)转录本,通过逆转录PCR(RT-PCR)扩增得到CA基因全长序列。生物信息学分析表明,FsCA1基因全长1 233 bp,开放阅读框978 bp,编码325个氨基酸;分子量35.11 ku,等电点7.59;包含9个α-螺旋、6个β-折叠、多个无规则卷曲和延伸链;含有1个信号肽和1个跨膜区;具有β-类碳酸酐酶典型的2个氨基酸保守域。亚细胞定位显示,该蛋白出现在叶绿体中的可能性最大。利用同源建模法构建了FsCA1三维结构模型,表明甜荞CA与豌豆CA同型八聚体能很好地匹配,推测甜荞CA也是同型八聚体。用实时荧光定量PCR检测FsCA1在荞麦不同器官中的表达水平,结果显示,FsCA1在叶中表达水平最高,其次是在茎中,在根中表达水平最低。 展开更多
关键词 西藏甜荞 碳酸酐酶 fsca1基因 三维结构预测 生物信息学
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Cloning and Bioinformation Analysis of Carbonic Anhydrase Gene FsCA1 of Tibet Wild Buckwheat
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作者 Weihai HOU Jianlin WANG +1 位作者 Danba Hudan 《Asian Agricultural Research》 2017年第11期54-57,65,共5页
A carbonic anhydrase( CA) transcript was obtained from the Contig library according to the published sequencing information of the buckwheat transcripts. The full length of the CA gene was amplified by reverse transcr... A carbonic anhydrase( CA) transcript was obtained from the Contig library according to the published sequencing information of the buckwheat transcripts. The full length of the CA gene was amplified by reverse transcription PCR( RT-PCR). The bioinformatics analysis showed that the full length of Fs CA1 gene was 1233 bp and open reading frame was 978 bp,and encoding 325 amino acids. The molecular weight was 35. 11 ku and the isoelectric point was 7. 59; there were 9 α helices,6 β folds,many randon coil and extension chain,containing one signal peptide and one transmembrane region,having a 2 amino acid conserved domains with typical beta-type carbonic anhydrase. Subcellular localization showed that the protein is most likely to appear in the chloroplast. The three-dimensional structure model of Fs CA1 was built by homologous modeling method,indicating that the homo-octamer of buckwheat CA and pea CA could match well,so it can be inferred that buckwheat CA is also homo-octamer. Real-time quantitative PCR was used to detect the expression of Fs CA1 in different organs of buckwheat.The results showed that Fs CA1 had the highest expression level in leaves,then in the stems,and the lowest in roots. 展开更多
关键词 Tibet buckwheat Carbonic anhydrase(CA) fsca1 Three-dimensional structure prediction BIOINFORMATICS
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