雷公藤PTEN基因全长克隆及表达分析  

Cloning and m RNA expression analysis of phosphatidylinositol-3,4,5-trisphosphate 3-phosphatase gene from Tripterygium wilfordii

作　　者：周家伟[1] 刘雨佳[1] 胡添源 高伟[1,2] 黄璐琦[3] 

机构地区：[1]首都医科大学中医药学院,北京100069 [2]首都医科大学中医络病研究北京市重点实验室,北京100069 [3]中国中医科学院中药资源中心,北京100700

出　　处：《中草药》2017年第7期1391-1396,共6页Chinese Traditional and Herbal Drugs

基　　金：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)

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 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.

关 键 词：雷公藤 磷脂酰肌醇-3 4 5-三磷酸肌醇3-磷酸酶 克隆 生物信息学分析 表达分析 

分 类 号：R282.12[医药卫生—中药学]