Cloning and Functional Analysis of a Vascular Tissue-specific Promoter from Populus tomentosa

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Cloning and Functional Analysis of a Vascular Tissue-specific Promoter from Populus tomentosa

Abstract

Promoter plays a key role not only in regulating the level of gene expression but also in controlling the expression in a spatial and temporal manner. According to the expression profile generated using Arabidopsis ATH1 genechip to analyze genes involved in the regeneration of the secondary vascular system in Populus tomentosa Carr, a differentially expressed gene NST3 was chosen and its homologous gene NAC068 in poplar were found using BLAST software against poplar EST database (PopulusDB). Then the 5′- upstream sequence of this gene was cloned from Populus tomentosa Carr. and put into the plant expression vector pBI121 to replace the 35S promoter to control the reporter gene gus. After transformation of polar 84K via Agrobactera, the GUS activity was detected in cambium zone, suggesting this promoter could control gene expression in secondary vascular system thus should be useful in genetic engineering to control exogenous gene specific expression in vascular system.
- vascular tissue,
- promoter,
- tissue-specific,
- GUS stain,
- genetic engineering

References

[1]	Plomion C, Leprovost G, Stokes A. Wood Formation in Trees[J]. Plant Physiology, 2001, 127(4): 1513-1523
[2] 卢孟柱, 胡建军. 我国转基因杨树的研究及应用现状[J]. 林业科技开发, 2006, 20(6): 1-4
[3] Kloti A, Henrich C, Bieri S, et al. Upstream and downstream sequence elements determine the specificity of the rice tungro bacilliform virus promoter and influence RNA production after transcription initiation[J]. Plant molecular biology, 1999, 40(2): 249-266
[4] Saito T, Tadakuma K, Takahashi N, et al. Two cytosolic cyclophilin genes of Arabidopsis thaliana differently regulated in temporal-and organ-specific expression[J]. Bioscience, biotechnology, and biochemistry, 1999, 63(4): 632-637
[5] Du J, Xie H L, Zhang D Q, et al. Regeneration of the secondary vascular system in poplar as a novel system to investigate gene expression by a proteomic approach[J]. Proteomics, 2006, 6(3): 881-895
[6] Wang M J, Qi X L, Zhao S T, et al. Dynamic changes in transcripts during regeneration of the secondary vascular system in Populus tomentosa Carr. revealed by cDNA microarrays[J]. BMC Genomics, 2009, 10: 215
[7] Murray M G, Thompson W F. Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Research, 1980, 8(19): 4321-4325
[8] Mitsuda N, Iwase A, Yamamoto H, et al. NAC transcription factors, NST1 and NST3, are key regulators of the formation of secondary walls in woody tissues of Arabidopsis[J]. The Plant Cell, 2007, 19(1): 270
[9] Zhong R, Richardson E A, Ye Z H. Two NAC domain transcription factors, SND1 and NST1, function redundantly in regulation of secondary wall synthesis in fibers of Arabidopsis[J]. Planta, 2007, 225(6): 1603-1611
[10]	[10] Mitsuda N, Seki M, Shinozaki K, et al. The NAC transcription factors NST1 and NST2 of Arabidopsis regulate secondary wall thickenings and are required for anther dehiscence[J]. The Plant Cell, 2005, 17(11): 2993-3006
[11]	[11] Zhong R, Demura T, Ye Z H. SND1, a NAC domain transcription factor, is a key regulator of secondary wall synthesis in fibers of Arabidopsis[J]. The Plant Cell, 2006, 18(11): 3158
[12]	[12] Sambrook J, Russell D W. Molecular cloning: a laboratory manual[M]. NY: Cold Spring Harbor Laboratory Press, 2001: 898-915
[13]	[13] Higo K, Ugawa Y, Iwamoto M, et al. Plant cis-acting regulatory DNA elements (PLACE) database[J]. Nucleic Acids Research, 1999, 27(1): 297-300
[14]	[14] Jefferson R A. Assaying chimeric genes in plants: the GUS gene fusion system[J]. Plant Molecular Biology Reporter, 1987, 5(4): 387-405
[15]	[15] Pedersen A G, Baldi P, Chauvin Y, et al. The biology of eukaryotic promoter prediction—a review[J]. Computers and Chemistry, 1999, 23(3-4): 191-207
[16]	[16] Lewin B. Gene VIII[M]. NJ: Pearson Prentice Hall, 2004
[17]	[17] Tjaden G, Coruzzi G M. A novel AT-rich DNA binding protein that combines an HMG I-like DNA binding domain with a putative transcription domain[J]. The Plant Cell, 1994, 6(1): 107-118
[18]	[18] Bustos M M, Guiltinan M J, Jordano J, et al. Regulation of β-Glucuronidase Expression in Transgenic Tobacco Plants by an A/T-Rich, cis-Acting Sequence Found Upstream of a French Bean β-Phaseolin Gene[J]. The Plant Cell, 1989, 1(9): 839-853
[19]	[19] Yang N S, Christou P. Cell type specific expression of a CaMV 35S-GUS gene in transgenic soybean plants[J]. Developmental Genetics, 1990, 11(4): 289-293
[20]	[20] Torres-schumann S, Ringli C, Heierli D, et al. In vitro binding of the tomato bZIP transcriptional activator VSF-1 to a regulatory element that controls xylem-specific gene expression.[J]. The Plant Journal, 1996, 9(3): 283
[21]	[21] Keller B, Baumgartner C. Vascular-specific expression of the bean GRP 1.8 gene is negatively regulated[J]. The Plant Cell, 1991, 3(10): 1051-1061
[22]	[22] Hatton D, Sablowski R, Yung M H, et al. Two classes of cis sequences contribute to tissue-specific expression of a PAL2 promoter in transgenic tobacco[J]. The Plant Journal, 1995, 7(6): 859-876
[23]

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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Cloning and Functional Analysis of a Vascular Tissue-specific Promoter from Populus tomentosa

Abstract: Promoter plays a key role not only in regulating the level of gene expression but also in controlling the expression in a spatial and temporal manner. According to the expression profile generated using Arabidopsis ATH1 genechip to analyze genes involved in the regeneration of the secondary vascular system in Populus tomentosa Carr, a differentially expressed gene NST3 was chosen and its homologous gene NAC068 in poplar were found using BLAST software against poplar EST database (PopulusDB). Then the 5′- upstream sequence of this gene was cloned from Populus tomentosa Carr. and put into the plant expression vector pBI121 to replace the 35S promoter to control the reporter gene gus. After transformation of polar 84K via Agrobactera, the GUS activity was detected in cambium zone, suggesting this promoter could control gene expression in secondary vascular system thus should be useful in genetic engineering to control exogenous gene specific expression in vascular system.

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Reference (23)

[1]	Plomion C, Leprovost G, Stokes A. Wood Formation in Trees[J]. Plant Physiology, 2001, 127(4): 1513-1523
[2]	[2] 卢孟柱, 胡建军. 我国转基因杨树的研究及应用现状[J]. 林业科技开发, 2006, 20(6): 1-4
[3]	[3] Kloti A, Henrich C, Bieri S, et al. Upstream and downstream sequence elements determine the specificity of the rice tungro bacilliform virus promoter and influence RNA production after transcription initiation[J]. Plant molecular biology, 1999, 40(2): 249-266
[4]	[4] Saito T, Tadakuma K, Takahashi N, et al. Two cytosolic cyclophilin genes of Arabidopsis thaliana differently regulated in temporal-and organ-specific expression[J]. Bioscience, biotechnology, and biochemistry, 1999, 63(4): 632-637
[5]	[5] Du J, Xie H L, Zhang D Q, et al. Regeneration of the secondary vascular system in poplar as a novel system to investigate gene expression by a proteomic approach[J]. Proteomics, 2006, 6(3): 881-895
[6]	[6] Wang M J, Qi X L, Zhao S T, et al. Dynamic changes in transcripts during regeneration of the secondary vascular system in Populus tomentosa Carr. revealed by cDNA microarrays[J]. BMC Genomics, 2009, 10: 215
[7]	[7] Murray M G, Thompson W F. Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Research, 1980, 8(19): 4321-4325
[8]	[8] Mitsuda N, Iwase A, Yamamoto H, et al. NAC transcription factors, NST1 and NST3, are key regulators of the formation of secondary walls in woody tissues of Arabidopsis[J]. The Plant Cell, 2007, 19(1): 270
[9]	[9] Zhong R, Richardson E A, Ye Z H. Two NAC domain transcription factors, SND1 and NST1, function redundantly in regulation of secondary wall synthesis in fibers of Arabidopsis[J]. Planta, 2007, 225(6): 1603-1611
[10]	[10] Mitsuda N, Seki M, Shinozaki K, et al. The NAC transcription factors NST1 and NST2 of Arabidopsis regulate secondary wall thickenings and are required for anther dehiscence[J]. The Plant Cell, 2005, 17(11): 2993-3006
[11]	[11] Zhong R, Demura T, Ye Z H. SND1, a NAC domain transcription factor, is a key regulator of secondary wall synthesis in fibers of Arabidopsis[J]. The Plant Cell, 2006, 18(11): 3158
[12]	[12] Sambrook J, Russell D W. Molecular cloning: a laboratory manual[M]. NY: Cold Spring Harbor Laboratory Press, 2001: 898-915
[13]	[13] Higo K, Ugawa Y, Iwamoto M, et al. Plant cis-acting regulatory DNA elements (PLACE) database[J]. Nucleic Acids Research, 1999, 27(1): 297-300
[14]	[14] Jefferson R A. Assaying chimeric genes in plants: the GUS gene fusion system[J]. Plant Molecular Biology Reporter, 1987, 5(4): 387-405
[15]	[15] Pedersen A G, Baldi P, Chauvin Y, et al. The biology of eukaryotic promoter prediction—a review[J]. Computers and Chemistry, 1999, 23(3-4): 191-207
[16]	[16] Lewin B. Gene VIII[M]. NJ: Pearson Prentice Hall, 2004
[17]	[17] Tjaden G, Coruzzi G M. A novel AT-rich DNA binding protein that combines an HMG I-like DNA binding domain with a putative transcription domain[J]. The Plant Cell, 1994, 6(1): 107-118
[18]	[18] Bustos M M, Guiltinan M J, Jordano J, et al. Regulation of β-Glucuronidase Expression in Transgenic Tobacco Plants by an A/T-Rich, cis-Acting Sequence Found Upstream of a French Bean β-Phaseolin Gene[J]. The Plant Cell, 1989, 1(9): 839-853
[19]	[19] Yang N S, Christou P. Cell type specific expression of a CaMV 35S-GUS gene in transgenic soybean plants[J]. Developmental Genetics, 1990, 11(4): 289-293
[20]	[20] Torres-schumann S, Ringli C, Heierli D, et al. In vitro binding of the tomato bZIP transcriptional activator VSF-1 to a regulatory element that controls xylem-specific gene expression.[J]. The Plant Journal, 1996, 9(3): 283
[21]	[21] Keller B, Baumgartner C. Vascular-specific expression of the bean GRP 1.8 gene is negatively regulated[J]. The Plant Cell, 1991, 3(10): 1051-1061
[22]	[22] Hatton D, Sablowski R, Yung M H, et al. Two classes of cis sequences contribute to tissue-specific expression of a PAL2 promoter in transgenic tobacco[J]. The Plant Journal, 1995, 7(6): 859-876
[23]

Cloning and Functional Analysis of a Vascular Tissue-specific Promoter from Populus tomentosa

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References

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通讯作者: 陈斌, bchen63@163.com

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Cloning and Functional Analysis of a Vascular Tissue-specific Promoter from Populus tomentosa

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