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排序方式: 共有1610条查询结果,搜索用时 31 毫秒
1.
Expression of cholera toxin B subunit oligomers in transgenic potato plants   总被引:36,自引:0,他引:36  
A gene encoding the cholera toxin B subunit protein (CTB), fused to an endoplasmic reticulum (ER) retention signal (SEKDEL) was inserted adjacent to the bi-directional mannopine synthase P2 promoter in a plant expression vector containing a bacterial luciferase AB fusion gene (luxF) linked to the P1 promoter. Potato leaf explants were transformed by Agrobacterium tumefaciens carrying the vector and kanamycin-resistant plants were regenerated. The CTB-SEKDEL fusion gene was identified in the genomic DNA of bioluminescent plants by polymerase chain reaction amplification. Immunoblot analysis indicated that plant-derived CTB protein was antigenically indistinguishable from bacterial CTB protein, and that oligomeric CTB molecules (Mr 50 kDa) were the dominant molecular species isolated from transgenic potato leaf and tuber tissues. Similar to bacterial CTB, plant-synthesized CTB dissociated into monomers (Mr 15 kDa) during heat or acid treatment. The maximum amount of CTB protein detected in auxin-induced transgenic potato leaf and tuber tissues was approximately 0.3% of total soluble plant protein. Enzyme-linked immunosorbent assay methods indicated that plant-synthesized CTB protein bound specifically to GM1-ganglioside, the natural membrane receptor of cholera toxin. In the presence of the SEKDEL signal, CTB protein accumulates in potato tissues and is assembled into an oligomeric form that retains native biochemical and immunological properties. The expression of oligomeric CTB protein with immunological and biochemical properties identical to native CTB protein in edible plants opens the way for preparation of inexpensive food plant-based oral vaccines for protection against cholera and other pathogens in endemic areas throughout the world  相似文献
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Summary Granule-bound starch synthase [GBSS; EC 24.1.21] determines the presence of amylose in reserve starches. Potato plants were transformed to produce antisense RNA from a gene construct containing a full-length granule-bound starch synthase cDNA in reverse orientation, fused between the cauliflower mosaic virus 35S promoter and the nopaline synthase terminator. The construct was integrated into the potato genome by Agrobacterium rhizogenes-mediated transformation. Inhibition of GBSS activity in potato tuber starch was found to vary from 70% to 100%. In those cases where total suppression of GBSS activity was found both GBSS protein and amylose were absent, giving rise to tubers containing amylose-free starch. The variable response of the transformed plants indicates that position effects on the integrated sequences might be important. The results clearly demonstrate that in tubers of potato plants which constitutively synthesize antisense RNA the starch composition is altered.  相似文献
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Russet Burbank potato plants have been genetically improved to resist insect attack and damage by Colorado potato beetles (Leptinotarsa decemlineata (Say)) by the insertion of a cryIIIA gene encoding the insect control protein of Bacillus thuringiensis var. tenebrionis. A modified gene that dramatically improved plant expression of this protein was utilized. Its expression in Russet Burbank potato plants resulted in protection from damage by all insect stages in the laboratory and in dramatic levels of protection at multiple field locations. Analysis of these genetically modified potatoes indicated that they conform to the standards for Russet Burbank potatoes in terms of agronomic and quality characteristics including taste.  相似文献
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Possible involvement of jasmonates in various morphogenic events   总被引:23,自引:0,他引:23  
Jasmonates (jasmonic acid and related compounds) seem to be involved in various morphogenic events of plants, such as tuberization (potato, yam and Jerusalem artichoke), tuberous root formation (sweet potato), bulb formation (onion and garlic), determination of plant structure (soybean) and thigmomorphogenesis (coiling of tendrils of Bryonia dioica ). The involvement of jasmonates in tuberization in these plants was inferred from their ability to induce tubers in vitro, and from changes in the levels of endogenous jasmonates during the growth of the plants, which can account for the initiation of tuberization. As to potato tuberization, jasmonic acid (JA) and its methyl ester (JA-Me) have strong tuber-inducing activity. These compounds seem to exert their tuber-inducing effects by elicting the expansion of cells, because JA and JA-Me are capable of causing the expansion of cells in potato tubers. The JA-induced expansion of cells is attributable to both an increase in osmotic pressure due to the accumulation of sucrose and changes in cell wall architecture that appear to affect the extensibility of the wall. And, moreover, the synthesis of cellulose might be indispensable for the JA-induced expansion. The tuberization and the expansion of cells induced by JA always involve the reorientation of cortical microtubules (MTs), suggesting that JA controls the direction of cell expansion by changing the arrangement of MTs. However, the reorientation of MTs itself seems to be insufficient for the induction of expansion of cells.
Involvement of jasmonates in bulb formation and tuberous root formation is presumed from the fact that JA is able to induce these in vitro. The exact nature of the control that the jasmonates exert on morphogenesis remains to be elucidated.  相似文献
7.
马铃薯块茎发育机理及其基因表达   总被引:23,自引:0,他引:23  
柳俊  谢从华 《植物学通报》2001,18(5):531-539
马铃薯(Solanum tuberosum L.)块茎是有块茎马铃薯植物的地下变态器官,它由匍匐茎顶端膨大形成,对于马铃薯块茎形成的生理机制已有许多研究,这些研究表明,块茎发生受许多因素的影响,总体来讲短日 照,较低的温度以及离体条件下培养基较高的蔗糖浓度等有利于块茎形成,同时,块茎形成过程中内源激素亦发生一系列变化,然而,对于块茎形成中相关基因表达,进而调控块茎形成的系统研究目前还较滞后,已有研究显示,块茎形成与膨大涉及到一系列基因的表达与关闭,同时它也与淀粉合成和块茎储藏蛋白基因的表达有关,综述了这一领域现有的研究进展。  相似文献
8.
高效马铃薯遗传转化体系的建立及甜蛋白基因的导入   总被引:22,自引:0,他引:22  
本研究选用了三个马铃薯(Solanum Iuberosum L.)栽培品种“85T-14-3”、“86-2”及“Favorita”的块茎、微型薯和试管薯为起始材料,应用根癌农杆菌 Ti 质粒系统成功地建立了一种方法简单、速度快和频率高的遗传转化体系。其中试管薯薄片的转化速度最快,经根癌农杆菌(Agrobacterium tumefaciens)共培养后,薄片在100mg/L 卡那霉素的分化培养上,2—3周就可产生出抗性小芽,这些小芽进一步仲长后可在50—100mg/L 卡那霉素的无激素MS 培养基上生根。从共培养到转化植株的获得只需6—7周。微型薯和试管薯的转化频率较高,最高可达67.5%。大多数抗性植株均能检测到胭脂碱合成酶或 GUS 基因的表达。把带有甜蛋白基因和胭脂碱合成酶标记基因的Ti质粒导入马铃薯,获得大量转化植株。叶片抗性检测和 nopaline 检测可推断外源甜蛋白基因已进入马铃薯基因组。  相似文献
9.
A cDNA fragment encoding human lactoferrin (hLF) linked to a plant microsomal retention signal peptide (SEKDEL) was stably integrated into the Solanum tuberosum genome by Agrobacterium tumefaciens-mediated leaf disk transformation methods. The lactoferrin gene was expressed under control of both the auxin-inducible manopine synthase (mas) P2 promoter and the cauliflower mosaic virus (CaMV) 35S tandem promoter. The presence of the hLF cDNA in the genome of regenerated transformed potato plants was detected by polymerase chain reaction amplification methods. Full-length hLF protein was identified by immunoblot analysis in tuber tissue extracts from the transformed plants by immunoblot analysis. The hLF produced in transgenic plant tissues migrated during polyacrylamide gel electrophoresis as a single band with an approximate molecular mass equal to hLF. Auxin activation of the mas P2 promoter increased lactoferrin expression levels in transformed tuber and leaf tissues to approximately 0.1% of total soluble plant protein. Antimicrobial activity against four different human pathogenic bacterial strains was detected in extracts of lactoferrin-containing potato tuber tissues. This is the first report of synthesis of full length, biologically active hLF in edible plants.  相似文献
10.
马铃薯叶片高效再生体系的建立   总被引:22,自引:0,他引:22  
以4个马铃薯栽培品种为试材,进行了叶片离体再生研究,结果表明:东农303、鄂1号叶片愈伤组织诱导的最佳培养基为MS+6-BA2.5mg·L-1+NAA0.2mg·L-1;费乌瑞它为MS+6-BA2.0mg·L-1+NAA0.1mg·L-1;夏波帝为MS+6-BA2.0mg·L-1+NAA0.2mg·L-1,愈伤组织的诱导率均可达100%.诱导不定芽分化的最佳培养基分别是:费乌瑞它、鄂1号为MS+6-BA2.5mg·L-1+GA35.0mg·L-1;东农303为MS+6-BA1.0mg·L-1+IAA0.1mg·L-1+GA32.5mg·L-1;夏波帝为MS+6-BA2.5mg·L-1+IAA0.5mg·L-1+GA32.5mg·L-1,其不定芽分化率分别达94.3%、100%、100%和90%.  相似文献
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