全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

棉花肉桂醇脱氢酶基因GhCAD6的克隆及正义、反义与RNAi干扰载体的构建

DOI: 10.7668/hbnxb.2009.06.005, PP. 20-26

Keywords: 棉花,肉桂醇脱氢酶,表达载体,RNAi

Full-Text   Cite this paper   Add to My Lib

Abstract:

肉桂醇脱氢酶(CAD)催化木质素单体合成的最后一步反应,将肉桂醛还原生成相应的肉桂醇.采用PCR方法从棉花中克隆了GhCAD6基因1569bp的基因组序列,序列分析表明GhCAD6基因由5个外显子和4个内含子组成.为了研究GhCAD6基因的功能,构建由棉纤维组织特异性启动子E6驱动的GhCAD6基因正义、反义表达载体;同时克隆了GhCAD6基因的一段41bp高度保守的NBD区序列,构建了GhCAD6基因的RNAi干扰载体,并将上述载体转入农杆菌菌株LBA4404中.为通过转基因技术深入研究GhCAD6基因在棉纤维发育和木质素代谢途径中的作用创造条件.

References

[1]  Fire A,Xu S Q,Montgomery M K,et al.Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans[J].Nature,1998,391(19):806-811.
[2]  Smith N A,Singh S P,Wang M B,et al.Total silencing by intron-spliced hairpin RNAs[J].Nature,2000,407:319-320.
[3]  Chuang C F,Meyerowitz E M.Specific and heritable genetic interference by double-stranded RNA in Arabidopsis thaliana[J].Proc Natl Acad Sci USA,2000,97:4985-4990.
[4]  康国章,肖向红,王永华,等.小麦淀粉GBSSH基因的克隆、反义和RNAi载体的构建[J].华北农学报,2008,23(1):7-11.
[5]  朱红林,沙爱华,符秀梅,等.转录调控基因GmLEC1的cDNA克隆及其植物表达载体的构建[J].华北农学报,2009,24(1):64-68.
[6]  Mansell R L,Gross G G,Stockigt J,et al.Purification and properties of cinnamyl alcohol dehydrogenase from higher plants involved in lignin biosynthesis[J].Phytochemistry,1974,13:2427-2435.
[7]  Halpin C,Knight M E,Foxon G A,et al.Manipulation of lignin quality by downregulation of cinnamyl alcohol dehydrogenase[J].Plant J,1994,6:339-350.
[8]  Baucher M,Chabbert B,Pilate G,et al.Red xylem and higher lignin extractability by down-regulating a cinnamyl alcohol dehydrogenase in poplar[J].Plant Physiol,1996,112:1479-1490.
[9]  Baucher M,Bernard-Vailhé M A,Chabbert B,et al.Down-regulation of cinnamyl alcohol dehydrogenase in transgenic alfalfa (Medicago sativa L.) and the effect on lignin composition and digestibility[J].Plant Mol Biol,1999,39:437-447.
[10]  Ralph J,MacKay J J,Hatfield R D,et al.Abnormal lignin in a loblolly pine mutant[J].Science,1997,277:235-239.
[11]  Fan L,Shi W J,Hu W R,et al.Molecular and biochemical evidence for phenylpropanoid synthesis and presence of wall-linked phenolics in cotton fibers[J].Journal of Integrative Plant Biology,received.
[12]  Paterson A H,Lan Der E S,Hewitt J D.A rapid method for extraction of cotton(Gossypium spp.) genomic DNA suitable for RFLP or PCR analysis[J].Plant Mol Biol Rep,1993,11:122-127.
[13]  Mette M F,Aufsatz W,van der Winden J,et al.Transcriptional silencing and promoter methylation triggered by double-stranded RNA[J].EMBO J,2000,19:5194-5520.
[14]  Sijen T,Vijn I,Rebocho A,et al.Transcriptional and posttranscriptional gene silencing are mechanistically related[J].Curr Biol,2001,11:436-444.
[15]  Chan S W-L,Zilberman D,Xie Z,et al.RNA silencing genes control de novo DNA methylation[J].Science,2004,303:1336.
[16]  黄冰艳,吉万全,郭蔼光,等.转录后基因沉默(PTGS)及其在作物遗传改良中的应用[J].中国生物工程杂志,2005,25(5):1-5.
[17]  Wesley S V,Helliwell C A,Smith N A,et al.Construct design for efficient,effective and high-throughput gene silencing in plants[J].Plant J,2001,27:581-590.
[18]  王雷,种康,许智宏.植物功能基因组学研究的有效工具-RNAi技术[J].植物生理学通讯,2003,39(6):705-710.
[19]  Neil A S,Surinder P S,Wang M B,et al.Total silencing by intron spliced hairpin RNAs[J].Nature,2001,407:319-320.
[20]  Hammond S M,Bernstern E,Beach D.An RNA-directed nuclease mediates post-transcriptional gene silencing in Drosophila cells[J].Nature,2000,404(16):293-296.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133