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作物学报 ›› 2011, Vol. 37 ›› Issue (04): 641-649.doi: 10.3724/SP.J.1006.2011.00641

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

三个桑树肌动蛋白基因的克隆与组织表达分析

李军,赵爱春**,王茜龄,张琼予,黎其友,金筱秐,李镇刚,余茂德*   

  1. 西南大学生物技术学院,重庆北碚400715
  • 收稿日期:2010-09-13 修回日期:2011-01-06 出版日期:2011-04-12 网络出版日期:2011-02-24
  • 基金资助:

    本研究由国家现代农业产业技术体系建设专项(nycytx-27-gw101)和重庆市蚕桑重大科技专项(CSTC, 2009AA1024)资助。

Molecular Cloning and Tissues Expression Analysis of Three Actin Genes from Mulberry (Morus alba)

LI Jun, ZHAO Ai-Chun**, WANG Xi-Lin, ZHANG Qiong-Yu, LI Qi-You, JIN Xiao-Yun, LI Zhen-Gang,YU Mao-De*   

  1. Collage of Biotechnology of Southwest University, Chongqing 400715, China
  • Received:2010-09-13 Revised:2011-01-06 Published:2011-04-12 Published online:2011-02-24

摘要: 肌动蛋白基因在植物各种生理活动中具有极其重要的作用,通过同源克隆与反向PCR的方法,克隆了3个肌动蛋白基因的核心片段,其中一个为已报道的MaACT1,另两个分别被命名为MaACT2MaACT3,进而PCR扩增获得MaACT1MaACT2肌动蛋白全长CDS,其中MaACT2基因全长1 704 bp,由4个外显子和3个内含子组成,CDS为1 134 bp,编码377个氨基酸残基。采用RT-PCR的方法分析了3个基因在叶、茎、果、根等组织的表达情况以及在茎、叶和托叶的生长过程中的表达变化。MaACT1在茎中表达量较弱但有随着茎的生长逐渐增强的趋势,在幼叶中有较高的表达,MaACT2MaACT3在根、茎、叶等组织中都有较高表达,MaACT3在叶、托叶和茎的各个发育时期表达都很稳定,可以作为桑树基因表达研究的内参基因。

关键词: 桑树, 肌动蛋白, 基因克隆, 表达分析, 进化

Abstract: Actins play many extremely important roles in plant, but none full-length cDNA actin genes were reported in mulberry until now. Three core fragments, two full CDS and one full-length genomic sequence of actin genes from mulberry were obtained with the strategies of homologous cloning and PCR in this research. Three mulberry actin genes obtained were designated as MaACT1, MaACT2, and MaACT3, respectively. The full-length of MaACT2 was 1 704 bp, which consisted of four exons and three introns, and 377 amino acidswere encoded by 1 134 bp of a putative CDS of MaACT2. The expression profile of three actins in leaf, stem, fruit and root was analyzed by RT-PCR. The results showed that MaACT1 was expressed low in stem, highen with the development of stem and high in young leaf. While MaACT2 and MaACT3 were expressed highly and stably in root, leaf and stem. The results showed MaACT3 was a good candidate for a control gene in the mulberry expression study because it can be expressed highly and stably in different tissues during their development.

Key words: Morus alba, Actin, Gene cloning, Expression analysis, Evolution

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