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中国农学通报 ›› 2017, Vol. 33 ›› Issue (12): 82-87.doi: 10.11924/j.issn.1000-6850.casb16050184

所属专题: 生物技术 园艺

• 林学 园艺 园林 • 上一篇    下一篇

苦瓜果实商品成熟期均一化全长cDNA文库的构建及分析

韩小霞,胡新军,李勇奇   

  1. 湖南省蔬菜工程技术研究中心/湖南省蔬菜研究所,湖南省蔬菜工程技术研究中心/湖南省蔬菜研究所,湖南省蔬菜工程技术研究中心/湖南省蔬菜研究所
  • 收稿日期:2016-05-31 修回日期:2017-04-12 接受日期:2016-08-25 出版日期:2017-04-28 发布日期:2017-04-28
  • 通讯作者: 韩小霞
  • 基金资助:
    湖南省自然科学基金“苦瓜果实商品成熟期均一化全长cDNA文库的构建及分析”(14JJ6050)。

Construction and Analysis of Normalized Full-length cDNA Library of Bitter Melon Fruit in Goods Mature Period

  • Received:2016-05-31 Revised:2017-04-12 Accepted:2016-08-25 Online:2017-04-28 Published:2017-04-28

摘要: 为构建苦瓜果实商品成熟期的cDNA文库,并对相关EST进行序列分析,笔者以多肽二号苦瓜亲本‘189-4’商品成熟期果实为材料,利用SMART技术构建其果实的cDNA文库。经检验该文库库容量为2.08×106,重组率为96.29%,平均插入片段750~2000 bp,插入片段平均大小约1000 bp。随机挑取400个单克隆进行测序,共获得370 条EST,序列分析结果表明350 条有同源性,全长率为70%;19 条序列无匹配,单一序列为320 条,其中,片段重叠群10 个,单基因260 个,可能与营养品质相关的基因有48 个。本研究为苦瓜果实功能基因的筛选、克隆奠定了基础。

关键词: 种植结构, 种植结构, 粮食生产, 演变, 影响因素, 研究回顾

Abstract: In order to construct a full-length cDNA library of bitter melon fruit in goods mature period and analyze the sequence of EST, in this research, we constructed a full-length cDNA library of bitter melon fruit for polypeptide bitter melon parent‘189-4’in goods mature period using the SMART technology. The tested results showed that the capacity of the library was 2.08×106, restructuring at a rate of 96.29%, the average insertion fragment ranged from 750 to 2000 bp, and the insert fragment average size was about 1000 bp. By sequenced 400 monoclonal randomly, we got 370 EST and the sequence analysis results showed that 350 had a homology, full length rate was 70%; 19 sequence had no matching, 320 was unigene, among them there were 10 contigs, 260 single genes, and there were about 48 genes possibly related to the nutrition quality. This research laid a foundation for the screening, cloning of functional genes for the bitter melon.

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