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目的:克隆红花Unigene100791基因,研究其表达情况;并对该基因的特性进行生物信息学分析,为后续红花代谢调控及提高红花油产量,使其满足食品工业市场需求奠定理论基础。方法:从红花转录物组数据库中筛选得到转录因子Dof相关基因Unigene100791的中间片段;利用RNAiso plus试剂法提取红花种子总RNA,利用RT-PCR方法反转录得到红花种子c DNA;采用3’Race方法从红花种子中克隆Unigene100791基因的全长序列;通过生物信息学方法对其全长基因进行分析,构建该基因与相关物种的系统进化树,研究其蛋白序列的同源性及其理化性质与结构特征,预测其生物学功能。结果:成功克隆了红花Unigene100791基因,其基因全长1274 bp;开放阅读框981 bp,编码326个氨基酸,理论分子质量约为35026.5 u,理论等电点9.5,带正电残基27个,带负电残基18个,序列含有典型的加尾信号poly(A);红花Unigene100791基因编码产物与烟草Dof2.4-1亲缘关系较近,属于Dof超家族,此蛋白存在信号肽,不存在跨膜结构。结论:成功克隆了红花Unigene100791基因,并对其编码的蛋白质进行了生物信息学分析。
OBJECTIVE: To clone the gene Unigene100791 of safflower, and to study the expression of Unigene100791. The bioinformatics analysis of this gene was carried out, which could lay the theoretical foundation for subsequent metabolism regulation of safflower and increase safflower oil production to meet the market demand of food industry. Methods: The middle fragment of Unigene100791, a transcription factor Dof gene, was screened from Safflower transcriptome database. The total RNA of Safflower seeds was extracted by RNAiso plus reagent and the cDNA of Safflower seeds was reverse transcribed by RT-PCR. 3’Race method was used to clone the full-length sequence of Unigene100791 from safflower seeds. The full-length gene of Unigene100791 was analyzed by bioinformatics method, and the phylogenetic tree of the gene and related species was constructed. Its physical and chemical properties and structural characteristics, predict its biological function. Results: The cDNA of Unigene100791 was successfully cloned. The full length cDNA of Unigene100791 was 1274 bp. The open reading frame was 981 bp encoding 326 amino acids. The theoretical molecular weight was about 35026.5 u, the theoretical isoelectric point was 9.5, There were 18 negatively charged residues with the sequence of poly (A). The gene of safflower Unigene100791 was closely related to tobacco Dof2.4-1 and belonged to the Dof superfamily. The signal peptide existed in this protein and did not exist Transmembrane structure. Conclusion: The Unigene100791 gene was successfully cloned and bioinformatics analysis of its encoded protein was carried out.