[关键词]
[摘要]
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cDNA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(MeJA)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。
[Key word]
[Abstract]
Objective To clone the full-length cDNA of phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase in Tripterygium wilfordii (TwPTEN) and predicted its biological functions. Methods The specific primers were designed based on the transcriptome data of Tripterygium wilfordii, TwPTEN gene was cloned by Polymerase Chain Reaction. The full-length cDNA of TwPTEN was then analyzed by a series of bioinformatics analysis. Results It was showed that the full length of TwPTEN cDNA was 2 247 bp encoding 614 amino acids. The theoretical isoelectric point was 5.84 and the molecular weight was about 66 900. Sequence alignment and phylogenetic analysis demonstrated that TwPTEN had relative close relationship to PTEN from other species. Differential expression analysis revealed that the relative expression level of TwPTEN increased significantly after being induced by methyl jasmonate (MeJA). It indicated that the TwPTEN gene was relative to the biosynthesis of secondary metabolites. Conclusion PTEN gene is firstly cloned from T. wilfordii, which provides a basis for further studying the functions of TwPTEN.
[中图分类号]
[基金项目]
国家自然科学基金面上项目(81373906);国家自然科学优秀青年科学基金项目(81422053)