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的克隆与分析

DOI: 10.7668/hbnxb.2010.02.005, PP. 23-29

Keywords: 棉花,肉桂醇脱氢酶,基因克隆,原核表达

Full-Text   Cite this paper   Add to My Lib

Abstract:

肉桂醇脱氢酶(CAD)是木质素生物合成过程中的一个关键酶类。根据棉花纤维特异表达cDNA文库分析得到的肉桂醇脱氢酶基因EST序列设计引物,采用RT-PCR技术从棉花中克隆了一个CAD基因,命名为GhCAD5(GenBank登录号为FJ848868)。研究结果表明:GhCAD5全长1488bp,具有一个1068bp的开放阅读框,5′非编码区为140bp,3′非编码区为280bp,编码355个氨基酸,预测分子量约为38.403kDa,等电点为7.67。氨基酸同源性分析发现,GhCAD5与拟南芥AtCAD1和水稻OsCAD4的同源性较高。利用PCR方法克隆了GhCAD5基因的基因组序列,长度为1695bp,包含6个外显子和5个内含子。将GhCAD5的ORF连接于原核表达载体pET-28a中,经IPTG诱导,获得了相应蛋白的表达。棉花GhCAD5基因的克隆为进一步研究其生物学功能和应用奠定了基础。

References

[1]  Whetten R,Sederoff R.Lignin biosynthesis[J].The Plant Cell,1995,7(7):1001-1013.
[2]  Boerjan W,Ralph J,Baucher M.Lignin biosynthesis[J].Annual Review of Plant Biology,2003,54(1):519-546.
[3]  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(11):2427-2435.
[4]  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(3):339-350.
[5]  Baucher M,Bernard-Vailhe 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(3):437-447.
[6]  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(4):1479-1490.
[7]  Ralph J,MacKay J J,Hatfield R D,et al.Abnormal lignin in a loblolly pine mutant[J].Science,1997,277(5323):235-239.
[8]  武耀廷,张恒木,刘进元.棉纤维细胞发育过程中纤维素的生物合成[J].棉花学报,2003,15(3):174-179.
[9]  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,2009,51(7):626-637.
[10]  王文生,王省芬,马峙英,等.棉花抗黄萎病相关基因筛选与亚克隆文库构建[J].华北农学报,2006,21(S1):147-150.
[11]  Wan C Y,Wilkins T A.A modified hot borate method significantly enhances the yield of high-quality RNA from cotton(Gossypium hirsutum L.)[J].Analytical Biochemistry,1994,223(1):7-12.
[12]  倪志勇,徐兆师,刘丽,等.小麦转录因子TaDREB6基因的克隆及鉴定[J].麦类作物学报,2008,28(3):357-363.
[13]  Ji S J,Lu Y C,Feng J X,et al.Isolation and analyses of genes preferentially expressed during early cotton fiber development by subtractive PCR and cDNA array[J].Nucleic Acids Res,2003,31(10):2534-2543.
[14]  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 Molecular Biology Reporter,1993,11(2):122-127.
[15]  魏建华,宋艳茹.木质素生物合成途径及调控的研究进展[J].植物学报,2001,43(8):771-779.
[16]  倪志勇,马文静,吕萌.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[J].华北农学报,2009,24(6):21-26.
[17]  刘宝业,梁国鲁,张增艳.水稻几丁质酶的原核表达、复性及体外抑菌活性的研究[J].核农学报,2009,23(3):369-374.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133