太子参肌动蛋白基因PhACT2的全长cDNA克隆与生物信息学分析

发布时间：2018-04-13 03:04

  本文选题：太子参 + 肌动蛋白　； 参考：《分子植物育种》2017年02期

【摘要】：根据太子参肌动蛋白基因PhACT2的已知片段设计特异引物,采用RACE技术扩增全长cDNA,并通过生物信息学软件对该基因进行结构分析。通过测序及序列拼接获得PhACT2全长cDNA序列,开放阅读框为1 134 bp,编码377个氨基酸残基,分子量为41.79 k D,理论等电点为5.30。与其他植物同源序列进行分析表明,其核苷酸序列与甜菜的同源性较高为88%,氨基酸序列的同源性均在95%以上。进化分析表明,PhACT2与拟南芥At ACT7的氨基酸序列仅存在4个特异位点的氨基酸差异。实时荧光定量PCR分析结果显示,PhACT2在太子参的不同器官、组织中具有稳定的表达,可作为内参基因。PhACT2全长cDNA序列的成功克隆和生物信息学分析为其在分子生物学领域的应用奠定基础。
[Abstract]:A specific primer was designed according to the known PhACT2 fragment of the actin gene of Pseudostellaria heterophylla. The full-length cDNA was amplified by RACE and the structure of the gene was analyzed by bioinformatics software.The full-length PhACT2 cDNA sequence was obtained by sequencing and sequence splicing. The open reading frame was 1 134 BP, encoding 377 amino acid residues with a molecular weight of 41.79 KD and a theoretical isoelectric point of 5.30.The results of homology analysis with other plants showed that the homology of nucleotide sequence with sugar beet was 888.The homology of amino acid sequence was more than 95%.Evolutionary analysis showed that there were only four amino acid differences in the amino acid sequence of ACT7 between Ph and Arabidopsis thaliana.The results of real-time fluorescence quantitative PCR analysis showed that PhACT2 was stably expressed in different organs and tissues of Pseudostellaria heterophylla.It can be used as the successful cloning and bioinformatics analysis of the full-length cDNA sequence of the internal reference gene .PhACT2, which lays the foundation for its application in the field of molecular biology.
【作者单位】： 贵阳中医学院;
【基金】：国家自然科学基金(81460579);国家自然科学基金(81160501) 贵州省研究生工作站建设项目(黔教研合JYSZ字[2014]016)共同资助
【分类号】：S567.53;Q943.2
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本文编号：1742621