三叶青扩展蛋白家族基因全长cDNA的克隆及其生物信息学和表达分析

发布时间：2018-03-22 04:15

本文选题：三叶青　切入点：扩展蛋白基因　出处：《中草药》2016年05期 　论文类型：期刊论文

【摘要】：目的克隆三叶青Tetrastigma hemsleyanum的扩展蛋白家族基因,对其进行器官表达分析,并初步探讨其功能。方法根据Gen Bank中登录的扩展蛋白基因保守区域序列设计1对简并引物,以三叶青球形块根c DNA为模板进行RT-PCR扩增,再结合RACE技术,获得三叶青扩展蛋白基因全长c DNA序列,命名为Th-exp。采用半定量RT-PCR和荧光定量PCR分析Th-exp基因在不同器官中的表达情况。结果克隆获得扩展蛋白家族基因,共782 bp,其中包含630 bp开放阅读框,编码209个氨基酸(Gen Bank登陆号KP693606)。Th-exp编码的蛋白质具有典型的扩展蛋白结构特征,N端含有8个半胱氨酸的结构域,C端含有4个保守的色氨酸结构域,中间含有组氨酸功能域。Blast分析显示,Th-exp与葡萄、醉蝶花中扩展蛋白基因具有相似性。半定量RT-PCR和荧光定量PCR分析表明,Th-exp在三叶青叶、茎、普通细根和球形块根中均有表达,但球形块根中表达强于其他器官。结论克隆得到三叶青扩展蛋白家族基因Th-exp全长c DNA,初步预测其参与了三叶青块根的发育过程。
[Abstract]:Objective to clone the extended protein family genes of three Ye Qing Tetrastigma hemsleyanum, analyze their organ expression, and explore their functions. Methods A pair of degenerate primers were designed according to the conserved region of the extension protein gene entered in Gen Bank. The full-length c DNA sequence of the c DNA gene of three Ye Qing globular tubers was obtained by RT-PCR amplification with RACE technique. The expression of Th-exp gene in different organs was analyzed by semi-quantitative RT-PCR and fluorescence quantitative PCR. The protein encoded by the KP693606).Th-exp encoding 209 amino acid sequence Gen Bank has typical structural characteristics of extended protein. The N-terminal domain containing 8 cysteine has four conserved tryptophan domains at the C-terminal. The intermediate histidine domain. Blast analysis showed that Th-exp gene was similar to that of grape. Semi-quantitative RT-PCR and fluorescence quantitative PCR analysis showed that Th-exp was expressed in green leaves, stems, fine roots and globular tubers. Conclusion the full-length Th-exp cDNA of the three Ye Qing extended protein family gene was obtained and it was preliminarily predicted that it was involved in the development of the root block of the three Ye Qing tubers.
【作者单位】：杭州师范大学生命与环境科学学院;
【基金】：浙江省药用植物种质改良与质量控制技术重点实验室开放项目(201302)
【分类号】：S567.2;Q943.2

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