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植物研究 ›› 2008, Vol. 28 ›› Issue (4): 408-411.doi: 10.7525/j.issn.1673-5102.2008.04.006

• 论文 • 上一篇    下一篇

甜菜M14品系花期cDNA文库的构建及特异表达基因的筛选

马春泉;张 莹;崔 颖;王 冰;王玉婷;刘金玲;王宏建;李海英*   

  1. (黑龙江大学生命科学学院分子生物学实验室,哈尔滨 150080)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2008-07-20 发布日期:2008-07-20
  • 通讯作者: 李海英
  • 基金资助:
     

Construction of cDNA Library in Florescence and Screening Specifically Expressed Genes in Sugar Beet M14

MA Chun-Quan;ZHANG Ying;CUI Ying;WANG Bing;WANG Yu-Ting;LIU Jin-Ling;WANG Hong-Jian;LI Hai-Ying*   

  1. (Bio-Molecular Lab of College of Life Sciences,Heilongjiang University,Harbin 150080)
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-07-20 Published:2008-07-20
  • Contact: LI Hai-Ying
  • Supported by:
     

摘要: 构建了甜菜M14品系在花期的ZAP表达载体的cDNA文库,利用抑制消减杂交方法所获得的M14品系特异表达的2个EST片段为探针筛选cDNA文库,获得了M14品系特异表达cDNA片段Me-86和Me-84;进一步利用RACE技术获得了基因M14-86 cDNA片段的全长序列。生物信息学分析表明M14品系特异表达基因M14-86的cDNA片段与cDNA片段Me-84分别与大花马齿苋(Portulaca grandiflora)及瓶子草(Sarracenia purpurea)的26S核糖体RNA具有很高的同源性。

关键词: cDNA文库, EST, 表达载体, 噬菌斑原位杂交, RACE技术

Abstract: The cDNA library of sugar beet M14 in florescence was constructed using ZAP expression vector. Two ESTs, which were specifically expressed in M14 and obtained by suppression subtractive hybridization, were used as probes to screen the cDNA library. Then, two cDNA fragments Me-84 and Me-86 specifically expressed in M14 were obtained. Full length cDNA sequence of gene M14-86 was later obtained using the method of RACE. Bioinformatic analysis of M14-86 and Me-84 cDNA fragments showed high homology with the 26S ribosomal RNA gene in Sarracenia purpurea and Portulaca grandiflora, respectively.

Key words: cDNA library, EST, expression vector, plaques in situ hybridization, method of RACE

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