杨树皮储藏蛋白基因启动子的克隆和功能研究*

摘要/Abstract

摘要：

杨树树皮储藏蛋白BSP是类似种子储藏蛋白的氮素储藏物,冬季在韧皮部薄壁细胞中大量积累,是落叶树氮代谢中的重要成分。为了研究BSP基因启动子在转基因植物中的表达特性,探索其在植物基因工程研究中潜在的应用价值,我们用PCR方法从美洲黑杨基因组中DNA扩增得到了BSA启动子片段。与GUS基因融合构建中间载体后,转化烟草,获得了一批PCR检测为阳性的转化再生植株。经GUS组织化学检测,发现若干转基因烟草的茎和叶柄韧皮部以及叶脉都呈GUS染色阳性,初步证明杨树BSP基因启动子确有韧皮部表达特性,可介导GUS基因在转基因烟草韧皮部特异表达。

关键词: 杨树, 韧皮部特异性启动子, GUS基因, 转基因烟草

Abstract:

Bark storage protein(BSP)of poplar, anitrogen reserve similar to seed storage protein, plays an important role in the nitrogen metabolism of deciduous trees.They present in large quantities in the phloem parenchyma cells in winter.In order to learn the expression pattern of BSP gene promoter in transgenic plants, and to explore its potential application in plant genetic engineering for phloem-specific expression of interested genes.BSP gene promoter fragment was amplified by PCR method from Populus deltoides genome DNA.Intermediate vector was constructed by inserting the promoter sequence upstream of GUS gene in pBI121, resulting in the replacement of the original CaMV 35S promoter by the BSP promoter. Regenerated tobacco plants obtained by transformation were analyzed by PCR to select the putative transgenic plans.Histochemical localization of GUS activity indicated that BSP promoter could confer a phloemspecific expression of GUS gene.

Key words: Populus deltoides, Phloem-specific promoter, GUS gene, Transgenic tobacco plants

蒋浩,秦红敏,田颖川,DeanW.Gabriel. 杨树皮储藏蛋白基因启动子的克隆和功能研究^*[J]. 林业科学, 1999, 35(5): 46-50.

Hao Jiang,Hongmin Qin,Yingchuan Tian,W.Gabriel Dean. CLONINGOF A PROMOTER FRAGMENTOF BARK STORAGE PROTEIN GENE FROM POPULUS DELTOIDES AND ITS FUNCTION IN TRANSGENIC TOBACCO PLANTS[J]. Scientia Silvae Sinicae, 1999, 35(5): 46-50.

图/表 5

图1

图2

图3

图4

图5

参考文献 0

	贾盘兴, 蔡金科, 等. 微生物遗传学实验技术. 科学出版社. 1992. 108- 113.
	Bustos MM , Guiltinan MJ , Jordano J , et al. Regulation of β- glucuronidase expression in trangenic tobacco plants by an A/T-rich, cis-acting sequence found upstream of a French bean β-phaseolin gene. The Plant Cell, 1989. 1, 839- 853.
	Castresana C , Garcia-Luque I , Alonso E , et al. Both positive and negative elements mediate expression of a photoregulated CAB gene from Nicotiana plumbaginofolia. EMBO J., 1988. 7, 1929- 1936. doi: 10.1002/j.1460-2075.1988.tb03030.x
	Coleman GD , Chen THH . Sequence of a poplar storage protein gene. Plant Physiology, 1993. 102, 1347- 1348. doi: 10.1104/pp.102.4.1347
	Czarnecka E , Nagao RT , Key JL , et al. Characterization of Gmhsp26-A, a stress gene encoding a divergent heat shock protein of soybean: heavy-metal-induced inhibition intron processing. Molecular Cell Biology, 1988. 8, 1113- 1122. doi: 10.1128/MCB.8.3.1113
	Jefferson RA . Assaying chimeric genes in plants: the GUS gene fusion system. Plant Molecular Biology Reporter, 1987. 5, 387- 405. doi: 10.1007/BF02667740
	Lam E , Kano - Murakami Y , Gilmartin P , et al. A metal-dependent DNA- binding protein interacts with a constitutive element of a light- responsive promoter. Plant Cell, 1990. 2, 857- 866.
	Riggs CD , Voelker TA , Chrispeels MJ . Cotyledon nuclear proteins bind to DNA fragments harboring regulatory elements of phytohem agglutinin genes. Plant Cell, 1989. 1, 609- 621. doi: 10.1677/jme.0.0020085
	Sambrook J, Fritsch E F, Maniatis T.1989. Molecular Cloning, Cold Spring Harbor Laboratory Press,
	Tjaden G , Coruzzi GM . A novel AT-rich DNA binding protein that combines an HMG I- like DNA binding domain with a putative transcription domain. The Plant Cell, 1994. 6, 107- 118.
	Yamamoto S , Nishihara M , Morikaw a H , et al. Promoter analysis of seed storage protein genes from Canavalia gladiata D.C.. Plant Molecular Biology, 1995. 27, 729- 741. doi: 10.1007/BF00020226

[1]	孙伟博,宫新栋,周燕,李红岩. 转玉米PEPC和PPDK基因杨树苗期的光合生理特性[J]. 林业科学, 2020, 56(7): 33-43.
[2]	何经纬,张伊莹,田呈明,熊典广,梁英梅. 区域景观格局对杨树锈病为害流行的影响——以北京延庆地区银白杨为例[J]. 林业科学, 2020, 56(4): 99-108.
[3]	刘文鑫,陈志成,代永欣,万贤崇. 水通道蛋白PIP1基因过表达杨树的光合生理过程对干旱和复水的响应[J]. 林业科学, 2020, 56(2): 69-78.
[4]	孙伟博,魏朝琼,马晓星,魏辉,诸葛强. 3类转基因南林895杨田间试验的安全性评估[J]. 林业科学, 2020, 56(10): 53-62.
[5]	张超, 王进茂, 赵洁, 庞丁玮, 张德健, 杨敏生. 转多基因欧美杨Bt基因表达特征[J]. 林业科学, 2019, 55(9): 61-70.
[6]	辛福梅, 闫小莉, 张长耀, 贾黎明. 西藏拉萨河谷区藏川杨和北京杨树干液流特征及其对环境因子的响应[J]. 林业科学, 2019, 55(2): 22-32.
[7]	张扬, 杜琳, 唐贤丰, 刘唤唤, 周功克, 柴国华. 杨树BAG基因的鉴定及表达模式分析[J]. 林业科学, 2019, 55(1): 138-145.
[8]	李建庆, 梅增霞, 杨忠岐. 不同林分杨树云斑天牛种群空间格局地统计学分析[J]. 林业科学, 2018, 54(3): 83-90.
[9]	张芹, 徐宗大, 赵凯, 李晓伟, 张罗沙, 张启翔. 梅花花青素苷调控基因PmMYB1的分离及功能分析[J]. 林业科学, 2018, 54(10): 64-72.
[10]	黄绢, 陈存, 张伟溪, 丁昌俊, 苏晓华, 黄秦军. 干旱胁迫对转JERF36银中杨苗木叶片解剖结构及光合特性的影响[J]. 林业科学, 2017, 53(5): 8-15.
[11]	尹吴, 孙伟博, 周燕, 诸葛强. 毛果杨Rubisco活化酶基因的克隆与功能分析[J]. 林业科学, 2017, 53(4): 83-95.
[12]	章小铃, 王留强, 孙佩, 吴立栓, 樊玮, 张进, 卢孟柱, 胡建军. 杨树不同种质花粉萌发和致敏蛋白的差异[J]. 林业科学, 2017, 53(2): 54-64.
[13]	李真, 王留强, 卢孟柱. 毛白杨PtoWOX11/12a对杨树扦插苗生长发育的影响[J]. 林业科学, 2017, 53(11): 69-76.
[14]	储文渊, 王玉娇, 朱东悦, 陈竹, 严涵薇, 项艳. 盐和干旱胁迫下杨树新内参基因的筛选[J]. 林业科学, 2017, 53(10): 70-79.
[15]	曹国玉, 李继祥, 买买提库尔班·阿力, 李开花, 胡建军. 杨树良种‘北杨’[J]. 林业科学, 2016, 52(9): 156-156.