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目的获得雷公藤Tripterygium wilfordii磷脂酰肌醇-3,4,5-三磷酸肌醇3-磷酸酶(Phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase,PTEN)基因的c DNA全长,并预测其生物学功能。方法根据雷公藤转录组数据设计引物,对雷公藤PTEN(TwPTEN)基因进行全长克隆;通过生物信息学方法对得到的TwPTEN基因进行分析,主要包括多重序列比对,蛋白结构预测和构建进化树分析等。结果根据分析结果 TwPTEN基因全长为2 247 bp,共编码614个氨基酸,等电点为5.84,相对分子质量为66 900,多重序列比对显示其与其他植物的PTEN基因具有较高的同源性。雷公藤悬浮细胞经外源茉莉酸甲酯(Me JA)诱导后,实时荧光定量结果显示,TwPTEN的表达量明显增加,并在12 h达到最高,提示PTEN基因与植物的次生代谢产物的产生有关。结论本研究首次从雷公藤悬浮细胞中克隆得到PTEN基因,为进一步研究TwPTEN基因的功能奠定基础。
OBJECTIVE: To obtain the full cDNA of Tripterygium wilfordii (Phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase) gene and to predict Its biological function. Methods According to the transcriptome data of Tripterygium wilfordii, primers were designed to clone the full-length PTEN (TwPTEN) gene. The TwPTEN gene was analyzed by bioinformatics methods, including multiple sequence alignment, protein structure prediction and construction of phylogenetic tree Analysis and so on. Results According to the results of the analysis, the full length of TwPTEN gene was 2 247 bp, encoding a total of 614 amino acids with an isoelectric point of 5.84 and a relative molecular mass of 66 900. Multiple sequence alignment showed that the gene of TwPTEN shared higher homology with other plant PTEN genes Sex. After induced by exogenous Me JA, the results of real-time fluorescence quantitative analysis showed that the expression of TwPTEN significantly increased and reached the highest level at 12 h, suggesting that the secondary metabolites of PTEN gene and plant related. Conclusions This study is the first to clone the PTEN gene from the suspension cells of Tripterygium solani and lay a foundation for further study on the function of TwPTEN gene.