|本期目录/Table of Contents|

[1]刘英华,赵桃,龙海,等.簇毛麦(Dasypyrum villosum) GA20ox基因的分离和鉴定[J].应用与环境生物学报,2009,15(04):479-482.[doi:10.3724/SP.J.1145.2009.00479]
 LIU Yinghua,ZHAO Tao,LONG Hai,et al.Cloning and Characterization of A cDNA Encoding Gibberellin 20-oxidase from Dasypyrum villosum[J].Chinese Journal of Applied & Environmental Biology,2009,15(04):479-482.[doi:10.3724/SP.J.1145.2009.00479]
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簇毛麦(Dasypyrum villosum) GA20ox基因的分离和鉴定()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
15卷
期数:
2009年04期
页码:
479-482
栏目:
研究论文
出版日期:
2009-08-25

文章信息/Info

Title:
Cloning and Characterization of A cDNA Encoding Gibberellin 20-oxidase from Dasypyrum villosum
作者:
刘英华赵桃龙海邓光兵潘志芬余懋群
中国科学院成都生物研究所 成都 610041
1西华大学 成都 610039;2上海工程技术大学 上海 201620
Author(s):
LIU YinghuaZHAO TaoLONG HaiDENG GuangbingPAN ZhifenYU Maoqun
1Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China
2Xihua University, Chengdu 610039, China; 3Shanghai University of Engineering Science, Shanghai 201620, China
关键词:
簇毛麦赤霉素GA20oxRT-PCR原核表达
Keywords:
Dasypyrum villosumgibberellinGA20oxRT-PCRheterologous expression
分类号:
S512.03 : Q78
DOI:
10.3724/SP.J.1145.2009.00479
文献标志码:
A
摘要:
通过RT-PCR技术克隆了簇毛麦(Dasypyrum villosum)赤霉素生物合成关键酶GA20ox基因的全长序列,命名为DvGA20ox,GenBank登录号为EU142949. 该基因全长1 080 bp,推测编码359个氨基酸的蛋白质,具有植物GA20ox基因的典型保守结构域LPWKET和NYYPXCQKP. 该基因推导编码蛋白质与小麦、大麦和黑麦草GA20ox蛋白的同源性分别为98%、97%和91%. 该序列重组到原核表达载体pET-32a(+)获得的重组子pET-32a(+)-DvGA20ox转化大肠杆菌BL21pLysS后,IPTG诱导表达,SDS-PAGE分析表明,DvGA20ox基因在大肠杆菌中获得了高效表达,融合蛋白相对分子质量为55×103,与理论值相符. 结果为深入研究DvGA20ox蛋白的结构与功能以及该基因与株高发育的关系奠定了基础. 图3 参19
Abstract:
Gibberellin 20oxidase (GA20ox) is a key regulatory enzyme in the GA-biosynthetic pathway of plants. The full-length cDNA of Dasypyrum villosum GA20ox (Designated as DvGA20ox) was isolated and consisted of 1 080-bp and encoded 359 amino acid residues. Comparative and bio-informatics analyses revealed that the deduced amino acid sequence of DvGA20ox shared 98%, 97% and 91% homology with those of wheat, barley and Lolium perenne, respectively, and contained conserved LPWKET and NYYPXCQKP domains. The amplified fragments were cloned into pET-32a (+) vector, and after restriction analysis the gene was transferred into Escherichia coli strain BL21. The recombinant gave rise to a 55×103 fusion protein in response to the IPTG induction. Fig 3, Ref 19

参考文献/References:

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备注/Memo

备注/Memo:
中国科学院成都生物研究所知识创新工程领域前沿项目(No. CIB-2007-LYQY-Q03)和中国科学院“西部之光”西部博士项目资助
更新日期/Last Update: 2009-08-26