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棉花学报 ›› 2011, Vol. 23 ›› Issue (1): 80-89.

• 研究与进展 • 上一篇    下一篇

海岛棉pepc基因的克隆及序列和表达分析(英文)

王德龙,于霁雯,喻树迅*,翟红红,范术丽,宋美珍,张金发   

  1. 中国农业科学院棉花研究所/农业部棉花遗传改良重点实验室,河南 安阳 455004
  • 收稿日期:2009-03-25 出版日期:2011-01-15 发布日期:2011-01-15
  • 通讯作者: yu@cricaas.com.cn
  • 作者简介:王德龙(1983-),男,硕士
  • 基金资助:

    国家十一五“863”计划优质高产棉花分子品种创制(2006AA100105)

Cloning, Sequence and Expression Analysis of Gossypium barbadense L. pepc Gene(in English)

WANG De-long, YU Ji-wen, YU Shu-xun*, ZHAI Hong-hong, FAN Shu-li, SONG Mei-zhen, ZHANG Jin-fa   

  1. Cotton Research Institute, Chinese Academy of Agriculture Sciences/Key Laboratory of Cotton Genetic Improvement, Ministry of Agriculture, Anyang, Henan 455004, China
  • Received:2009-03-25 Online:2011-01-15 Published:2011-01-15
  • Contact: yu@cricaas.com.cn

摘要: 磷酸烯醇式丙酮酸羧化酶 (Phosphoenolpyruvate carboxylase)对植物生理功能行使重要的调节作用。本研究根据NCBI已公布的EST序列利用RACE和genome walking技术从海岛棉品种7124中克隆了一个新的pepc基因,命名为Gb.pepc3。该序列全长3259 bp,含有一个2910 bp的开放阅读框,编码969个氨基酸,推测分子量为110.7 kd,等电点为6.08 I。对Gb.pepc3蛋白的同源性比对和系统进化树分析显示,Gb.pepc3与已报道的其他植物的pepc相似性很高,属于C3pepc。荧光定量PCR结果显示Gb.pepc3在棉花各组织中广泛存在,其中在胚中表达量最高,在纤维中表达量较低。在棉花发育各个阶段Gb.pepc3表达量不同,海岛棉在开花后15天Gb.pepc3表达量达到高峰期,而陆地棉开花后20天Gb.pepc3表达量达到高峰期。海岛棉和陆地棉间表达量差异明显。

关键词: pepc; 克隆; 差异表达; 定量PCR

Abstract: According to published EST sequences, an new pepc(Phosphoenolpyruvate carboxylase) gene was cloned from Gossypium barbadense L. cultivar 7124 by the utilization of the RACE and genome walking technology. The gene was named Gb.pepc3. It consists of 3259 bps, contains an open reading frame for 2910 bps, and encodes 969 amino acids with a deduced molecular weight of 110.7 kd and an isoelectric point of 6.08 I. Homology and phylogenetic trees analysis showed high similarity between Gb.pepc3 and pepcs in other reported plants, and all belong to C3 type pepc. The result of the fluorescence quantitative PCR displayed that Gb.pepc3 existed in all tissues of cotton. It expressed highly in the embryo but low in fiber. The expression quantity of Gb.pepc3 varied in different stages of the development of cotton. The expression of Gb.pepc3 reached the highest level at 15 days after blossoming for Gossypium barbadense L. while 20 days after blossoming for G. hirsutum, and there was significant difference in the expression quantities between two species.

Key words: pepc; clone; differential expression; quantitative PCR; cotton

中图分类号: 
  • Q78: S562