Cloning and Sequence Analysis of 1-Deoxy-D-Xylulose 5-Phosphate Gene cDNA from Eucommia ulmoides

LIU Pan-feng; DU Hong-yan; WUYUN Ta-na; HUANG Hai-yan; ZHU Gao-pu

Volume 25 Issue 2

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Cloning and Sequence Analysis of 1-Deoxy-D-Xylulose 5-Phosphate Gene cDNA from Eucommia ulmoides

1.
Non-timber Forestry Research and Development Center, Chinese Academy of Forestry, Zhengzhou 450003, He’nan, China
2.
Key Laboratory of Non-wood Forest Products of State Forestry Administration, Changsha 410004 Hu’nan, China

Received Date: 2011-11-06

Abstract

Homologous DXR gene cDNA was isolated from leaves of Eucommia ulmoides by method of reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) technique, and named after EuDXR. The nucleotide sequence of EuDXR was 1 814 bp and contains an open reading frame (ORF) of 1 437 bp encoding 478 amino acids with a molecular weight of about 51.71 kD,and the theoretical isoelectric point of which was 5.79, representative motifs of plant DXR and 26 potential functional sites were deduced in the amino acids sequence of EuDXR. The results of phylogenetic analysis suggested that the protein sequence of EuDXR was more similar to that of Zea mays and Oryza sativa than other species, and followed by Hevea brasiliensis, Arabidopsis thaliana and Nicotiana tabacum.
- Eucommia ulmoides,
- DXR,
- gene,
- sequence analysis

References

[1]	Bohlmann J, Keeling C I. Terpenoid biomaterials[J]. Plant J, 2008,54(4): 656-669
[2]	Page J E, Hause G, Raschke M, et al. Functional analysis of the final steps of the 1-deoxy-D-xylulose-5-phosphate (DXP) pathway to isoprenoids in plants using virus-induced gene silencing[J]. Plant Physiol, 2004, 134: 1401-1413
[3]	Buchanan B B, 等.植物生物化学与分子生物学[M]. 瞿礼嘉,等译. 北京:科学出版社, 2004: 1251-1253
[4]	McGarvey D J, Croteau R. Terpenoid metabolism[J]. Plant Cell, 1995, 7: 1015-1026
[5]	Rohmer M, Knani M, Simonin P, et al. Isoprenoid biosynthesis in bacteria: a novel pathway for the early steps leading to isopentenyl diphophate[J]. Biochem J, 1993, 295: 517-524
[6]	马靓,丁鹏,杨广笑,等. 植物类萜生物合成途径及关键酶的研究进展[J]. 生物技术通报,2006(增刊):22-30
[7]	Mahmoud S S, Croteau R B. Metabolic engineering of essential oil yield and composition in mint by altering expression of deoxyxylilose phosphate reductoisomeras and menthofuran synthase[J]. Proc Natl Acad Sci, 2001, 98: 8915-8920
[8]	Carretero-Paulet L, Cair A, Botella-Pavia P, et al. Enhanced flux the methylerythritol 4-phosphate pathway in Arabidopsis plants overexpressing 1-deoxy-D-xylulose 5-phosphate reductoisomerase[J]. Plant Mol Biol, 2006, 62: 683-695
[9]	Tomohisa H, Shinya T, Shunsuke H, et al. Overexpression of 1-deoxy-D-xylulose-5-phosphate reductoisomerase gene in chloroplast contributes to increment of isoprenoid production[J]. J Bioscience Bioeng, 2008, 105: 518-526
[10]	Rodriguez-Concepcion M, Ahumada I, Diez-Juez E, et al. 1-deoxy-D-xylulose-5-phosphate reductoisomerase and plastid isoprenoid biosynthesis during tomato fruit ripening[J]. Plant J, 2001, 27: 213
[11]	Khemvong S, Suvachittanont W. Molecular cloning and expression of a cDNA encoding 1-deoxy-D-xylulose-5-phosphate synthase from oil palm Elaeis guineensis Jacq[J].Plant Sci,2005,169:571-578
[12]	Yortyot S,Thomas D,Wallie S. Molecular cloning and characterization of two cDNAs encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Hevea brasiliensis[J]. J Plant Physiol, 2008, 165: 991-1002
[13]	李芳东,杜红岩. 杜仲[M]. 北京:中国中医药出版社,2001
[14]	杜红岩. 杜仲活性成分与药理研究的新进展[J]. 经济林研究,2003,21(2):58-61
[15]	严瑞芳. 杜仲橡胶的开发及应用概况[J]. 橡胶科技市场,2010(10):9-13
[16]	陈建. 几种提取杜仲RNA方法的比较[J]. 林业科技开发,2007,21(5):19-21
[17]	周明兵,王红珍,赵德刚. 杜仲叶和树皮总RNA的快速提取法提取法[[J]. 山地农业生物学报,2003,22(5):430-431
[18]	Henrik N, Jacob E, Sren B, et al. Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites[J]. Protein Eng, 1997, 10:1-6
[19]	Arnold K, Bordoli L, Kopp J, et al. The SWISS-MODEL Workspace: A web-based environment for protein structure homology modelling[J]. Bioinformatics, 2006, 22: 195-201
[20]	Tamura K, Peterson D, Peterson N, et al. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Mol Biol Evol, 2011, 28: 2731-2739
[21]	Hongyan Y, Yifu G, Kaijing Z, et al. Molecular cloning, expression profiling and functional analysis of a DXR gene encoding 1-deoxy- D -xylulose 5-phosphate reductoisomerase from Camptotheca acuminate[J]. J Plant Physiol, 2008, 165: 203-213
[22]	Krueawan Y, Seiji T, Atiya R, et al. cDNA, from Hevea brasiliensis latex, encoding 1-deoxy- D -xylulose-5-phosphate reductoisomerase[J]. Plant Sci, 2008, 175: 694-700
[23]	Lorenzo C P, Iván A, Nuria C, et al. Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-deoxy-d-xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway[J]. Plant Physiol, 2002, 129: 1581-1591
[24]	von Heijne G, Stepphuhn J, Herrmann R G. Domain structure of mitochondrial and chloroplast targeting peptides[J]. Eur J Biochem, 1989, 180: 535-545

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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Cloning and Sequence Analysis of 1-Deoxy-D-Xylulose 5-Phosphate Gene cDNA from Eucommia ulmoides

1. Non-timber Forestry Research and Development Center, Chinese Academy of Forestry, Zhengzhou 450003, He’nan, China

2. Key Laboratory of Non-wood Forest Products of State Forestry Administration, Changsha 410004 Hu’nan, China

Received Date: 2011-11-06

Available Online: 2012-04-13

Abstract: Homologous DXR gene cDNA was isolated from leaves of Eucommia ulmoides by method of reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) technique, and named after EuDXR. The nucleotide sequence of EuDXR was 1 814 bp and contains an open reading frame (ORF) of 1 437 bp encoding 478 amino acids with a molecular weight of about 51.71 kD,and the theoretical isoelectric point of which was 5.79, representative motifs of plant DXR and 26 potential functional sites were deduced in the amino acids sequence of EuDXR. The results of phylogenetic analysis suggested that the protein sequence of EuDXR was more similar to that of Zea mays and Oryza sativa than other species, and followed by Hevea brasiliensis, Arabidopsis thaliana and Nicotiana tabacum.

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Reference (24)

[1]	Bohlmann J, Keeling C I. Terpenoid biomaterials[J]. Plant J, 2008,54(4): 656-669
[2]	Page J E, Hause G, Raschke M, et al. Functional analysis of the final steps of the 1-deoxy-D-xylulose-5-phosphate (DXP) pathway to isoprenoids in plants using virus-induced gene silencing[J]. Plant Physiol, 2004, 134: 1401-1413
[3]	Buchanan B B, 等.植物生物化学与分子生物学[M]. 瞿礼嘉,等译. 北京:科学出版社, 2004: 1251-1253
[4]	McGarvey D J, Croteau R. Terpenoid metabolism[J]. Plant Cell, 1995, 7: 1015-1026
[5]	Rohmer M, Knani M, Simonin P, et al. Isoprenoid biosynthesis in bacteria: a novel pathway for the early steps leading to isopentenyl diphophate[J]. Biochem J, 1993, 295: 517-524
[6]	马靓,丁鹏,杨广笑,等. 植物类萜生物合成途径及关键酶的研究进展[J]. 生物技术通报,2006(增刊):22-30
[7]	Mahmoud S S, Croteau R B. Metabolic engineering of essential oil yield and composition in mint by altering expression of deoxyxylilose phosphate reductoisomeras and menthofuran synthase[J]. Proc Natl Acad Sci, 2001, 98: 8915-8920
[8]	Carretero-Paulet L, Cair A, Botella-Pavia P, et al. Enhanced flux the methylerythritol 4-phosphate pathway in Arabidopsis plants overexpressing 1-deoxy-D-xylulose 5-phosphate reductoisomerase[J]. Plant Mol Biol, 2006, 62: 683-695
[9]	Tomohisa H, Shinya T, Shunsuke H, et al. Overexpression of 1-deoxy-D-xylulose-5-phosphate reductoisomerase gene in chloroplast contributes to increment of isoprenoid production[J]. J Bioscience Bioeng, 2008, 105: 518-526
[10]	Rodriguez-Concepcion M, Ahumada I, Diez-Juez E, et al. 1-deoxy-D-xylulose-5-phosphate reductoisomerase and plastid isoprenoid biosynthesis during tomato fruit ripening[J]. Plant J, 2001, 27: 213
[11]	Khemvong S, Suvachittanont W. Molecular cloning and expression of a cDNA encoding 1-deoxy-D-xylulose-5-phosphate synthase from oil palm Elaeis guineensis Jacq[J].Plant Sci,2005,169:571-578
[12]	Yortyot S,Thomas D,Wallie S. Molecular cloning and characterization of two cDNAs encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Hevea brasiliensis[J]. J Plant Physiol, 2008, 165: 991-1002
[13]	李芳东,杜红岩. 杜仲[M]. 北京:中国中医药出版社,2001
[14]	杜红岩. 杜仲活性成分与药理研究的新进展[J]. 经济林研究,2003,21(2):58-61
[15]	严瑞芳. 杜仲橡胶的开发及应用概况[J]. 橡胶科技市场,2010(10):9-13
[16]	陈建. 几种提取杜仲RNA方法的比较[J]. 林业科技开发,2007,21(5):19-21
[17]	周明兵,王红珍,赵德刚. 杜仲叶和树皮总RNA的快速提取法提取法[[J]. 山地农业生物学报,2003,22(5):430-431
[18]	Henrik N, Jacob E, Sren B, et al. Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites[J]. Protein Eng, 1997, 10:1-6
[19]	Arnold K, Bordoli L, Kopp J, et al. The SWISS-MODEL Workspace: A web-based environment for protein structure homology modelling[J]. Bioinformatics, 2006, 22: 195-201
[20]	Tamura K, Peterson D, Peterson N, et al. MEGA5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods[J]. Mol Biol Evol, 2011, 28: 2731-2739
[21]	Hongyan Y, Yifu G, Kaijing Z, et al. Molecular cloning, expression profiling and functional analysis of a DXR gene encoding 1-deoxy- D -xylulose 5-phosphate reductoisomerase from Camptotheca acuminate[J]. J Plant Physiol, 2008, 165: 203-213
[22]	Krueawan Y, Seiji T, Atiya R, et al. cDNA, from Hevea brasiliensis latex, encoding 1-deoxy- D -xylulose-5-phosphate reductoisomerase[J]. Plant Sci, 2008, 175: 694-700
[23]	Lorenzo C P, Iván A, Nuria C, et al. Expression and molecular analysis of the Arabidopsis DXR gene encoding 1-deoxy-d-xylulose 5-phosphate reductoisomerase, the first committed enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway[J]. Plant Physiol, 2002, 129: 1581-1591
[24]	von Heijne G, Stepphuhn J, Herrmann R G. Domain structure of mitochondrial and chloroplast targeting peptides[J]. Eur J Biochem, 1989, 180: 535-545

Cloning and Sequence Analysis of 1-Deoxy-D-Xylulose 5-Phosphate Gene cDNA from Eucommia ulmoides

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通讯作者: 陈斌, bchen63@163.com

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Cloning and Sequence Analysis of 1-Deoxy-D-Xylulose 5-Phosphate Gene cDNA from Eucommia ulmoides

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