生物技术通报 ›› 2015, Vol. 31 ›› Issue (5): 106-112.doi: 10.13560/j.cnki.biotech.bull.1985.2015.05.017

• 研究报告 • 上一篇    下一篇

沙冬青脯氨酸转运体基因的克隆及表达分析

岳光振, 金曼, 李俊林, 杨顺瑛, 郭满园, 苏彦华,   

  1. (中国科学院南京土壤研究所 土壤与农业可持续发展国家重点实验室,南京 210008)
  • 收稿日期:2014-09-11 出版日期:2015-05-18 发布日期:2015-05-18
  • 作者简介:岳光振,男,硕士研究生,研究方向:植物逆境生理生化特征;E-mail:biboqingyan@163.com
  • 基金资助:
    国家自然科学基金重大研究计划项目(91125028)

Cloning and Expression Analysis of Proline Transporter Gene in Ammopiptanthus mongolicus

Yue Guangzhen, Jin Man, Li Junlin, Yang Shunying, Guo Manyuan, Su Yanhua   

  1. (State Key Laboratory of Soil and Sustainable Agriculture/Institute of Soil Science,Chinese Academy of Sciences,Nanjing 210008)
  • Received:2014-09-11 Published:2015-05-18 Online:2015-05-18

摘要: 通过RACE技术从沙冬青中克隆获得了脯氨酸转运体基因AmProT(GenBank登录号为KJ873133)。序列分析表明,AmProT基因开放阅读框为1 329 bp,编码442个氨基酸,预测蛋白质分子量为48.07 kD,等电点为9.32,含有11个跨膜区域,具有典型的脯氨酸转运蛋白的特征。进化分析显示,AmProT与大豆ProT的相似性达到86%。荧光定量PCR分析表明,AmProT在地上部的表达量要显著高于地下部,在受到干旱、高盐、脱落酸胁迫后表达量呈上调趋势,推测AmProT可能在沙冬青响应干旱、盐等非生物胁迫过程中发挥作用。

关键词: 沙冬青, 脯氨酸转运体基因, 基因克隆, 表达分析

Abstract: The cDNA of proline transporter gene in Ammopiptanthus mongolicus(GenBank accession No. KJ873133)was amplified by RACE technology. Bioinformatics analysis showed that the length of AmProT ORF was 1 329 bp, the gene encoded a putative polypeptide of 442 amino acids with a calculated molecular mass of 48.0733 kD and a theoretical pI of 9.32. It had 11 transmembrance domains, which is the typical characteristics of proline translocator. Phylogenic analysis indicated that AmProT was 86% similar to proline transporter gene of Glycine max. Real-time PCR analysis revealed that AmProT expressed in all tissues examined with more higher level in the above-ground part than the underground part; expression profiles under different stress treatments such as drought, salt and ABA were compared, and the results revealed that transcriptional level of AmProT was up-regulated. It is speculated that AmProT plays an important role in response to abiotic stresses such as drought and salt.

Key words: Ammopiptanthus mongolicus, proline transporter gene, gene cloning, expression analysis