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拟南芥冷诱导型启动子CBF 3的克隆及活性检测 总被引:1,自引:0,他引:1
目的:构建冷诱导型启动子CBF3基因的植物表达载体,并将其转入烟草。方法:以拟南芥基因组DNA为模板,通过特异PCR扩增,克隆冷诱导表达启动子CBF3(C-repeat binding factor)。用CBF3启动子替换pBI121载体上的35S启动子构建新的载体pBC-GUS,通过农杆菌介导的叶盘法转化烟草。结果:获得了转基因烟草,转基因烟草的GUS组织化学染色及PCR分析结果表明,在低温诱导下,CBF3启动子可增强GUS基因表达。结论:CBF3启动子可应用于植物抗冷基因工程研究。 相似文献
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生长素结合蛋白cDNA的克隆及其在烟草中的表达 总被引:3,自引:0,他引:3
基于拟南芥内质网生长素结合蛋白基因的cDNA序列,设计合成了Ap5和Ap3两个引物,应用RT-PCR技术扩增了拟南芥的ABP基因。将该基因克隆在植物表达载体p35SSIN的35S启动子和Nos3’端之间,得到植物表达载体p35SE。通过农杆菌个导的方法对烟草SR1进行了转化,由分子杂交等检测证明,生长素结合蛋白基因已在烟草中表达,同时转基因烟草后代对生长素的敏感性明显增加。 相似文献
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《基因组学与应用生物学》2016,(11)
本研究利用PCR技术从’赤霞珠’葡萄(Vitis vinifera L.cv Cabernet Sauvignon)基因组DNA中扩增得到与花色素3,5-O-双葡萄糖苷合成相关的Vv5GT3基因的启动子。对启动子序列的顺式作用元件进行预测分析发现,Vv5GT3基因的启动子序列除了含有植物启动子的基本结构元件外,还有多个与胁迫和激素诱导相关的顺势调控元件。以荧光素酶基因(Luc)作为为报告基因,构建了Vv5GT3基因启动子的植物表达载体p CAMBIA1300-Vv5GT3 promoter。以根癌农杆菌GV3101感受态细胞为受体,转化植物融合表达载体进行瞬时表达,并对启动子进行了功能活性分析,结果表明该启动子能够驱动Luc报告基因在烟草叶片组织中表达。用不同的光照条件处理被农杆菌侵染的烟草叶片,结果显示长时间光照有利于启动子的激活,而遮光减弱了Vv5GT3启动子的活性。 相似文献
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甘蓝型油菜BcNA1基因启动子在转基因烟草中对GUS基因表达的调控 总被引:10,自引:0,他引:10
通过PCR扩增,从甘蓝型油菜(Brassica napus)品种H165中克隆了种子贮藏蛋白基因BcNAl的启动子,将此启动子与GUS基因相连构建了植物表达载体,利用农杆菌介导法将其导入烟草,对转基因烟划GUS基因检测分析表明,BcNAl基因启动子能特异地启动GUS基因在种子中的表达;而且GUS酶的活性随着转基因烟草种子的发育而变化。同时还间接证明BcNA1基因的启动子在转基因烟草中的遗传传递方式符合盂德尔遗传定律。 相似文献
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六棱大麦HVA1基因在烟草中遗传转化的研究 总被引:2,自引:0,他引:2
本研究依据HVA1基因序列克隆六棱大麦HVA1基因cDNA片段,构建Ubiquitin启动子驱动下的植物表达载体pCAMBIA1300-HVA1。然后通过三亲杂交法将重组质粒PCAMBIA1300-HVA1转入农杆菌LBA4404,并采用农杆菌介导法转化烟草。经PCR,PCR-Southern blotting和RT-PCR检测表明HVA1基因已整合进烟草基因组,并在转录水平上获得表达。功能验证的结果显示,转基因植株叶片的保水率提高了近1倍,暗示转基因烟草具有一定的抗旱潜力。 相似文献
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中南民族大学生命科学学院李静、刘学群和王春台3位科研工作者利用从烟草中克隆的一个葡糖基转移酶基因(GT—like),通过PCR扩增得到该基因开放阅读框5’端-1150~0上游调控区,经序列分析表明该启动子序列含有多个基因表达调控元件。将GT—Like基因启动子与gus报告基因连接,构建植物表达载体pGT-gus,经根癌农杆菌介导法转化W38型烟草。转基因烟草植株GUS组织化学染色结果表明: 相似文献
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In experiments on Black Sea skates (Raja clavata), the potential of the receptor epithelium of the ampullae of Lorenzini and spike activity of single nerve fibers connected to them were investigated during electrical and temperature stimulation. Usually the potential within the canal was between 0 and –2 mV, and the input resistance of the ampulla 250–400 k. Heating of the region of the receptor epithelium was accompanied by a negative wave of potential, an increase in input resistance, and inhibition of spike activity. With worsening of the animal's condition the transepithelial potential became positive (up to +10 mV) but the input resistance of the ampulla during stimulation with a positive current was nonlinear in some cases: a regenerative spike of positive polarity appeared in the channel. During heating, the spike response was sometimes reversed in sign. It is suggested that fluctuations of the transepithelial potential and spike responses to temperature stimulation reflect changes in the potential difference on the basal membrane of the receptor cells, which is described by a relationship of the Nernst's or Goldman's equation type.I. P. Pavlov Institute of Physiology, Academy of Sciences of the USSR, Leningrad. I. M. Sechenov, Institute of Evolutionary Physiology and Biochemistry, Academy of Sciences of the USSR, Leningrad. Pacific Institute of Oceanology, Far Eastern Scientific Center, Academy of Sciences of the USSR, Vladivostok. Translated from Neirofiziologiya, Vol. 12, No. 1, pp. 67–74, January–February, 1980. 相似文献
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N. P. Vesselkin Yu. V. Natochin 《Journal of Evolutionary Biochemistry and Physiology》2010,46(6):592-603
Evolution of living organisms is closely connected with evolution of structure of the system of regulations and its mechanisms.
The functional ground of regulations is chemical signalization. As early as in unicellular organisms there is a set of signal
mechanisms providing their life activity and orientation in space and time. Subsequent evolution of ways of chemical signalization
followed the way of development of delivery pathways of chemical signal and development of mechanisms of its regulation. The
mechanism of chemical regulation of the signal interaction is discussed by the example of the specialized system of transduction
of signal from neuron to neuron, of effect of hormone on the epithelial cell and modulation of this effect. These mechanisms
are considered as the most important ways of the fine and precise adaptation of chemical signalization underlying functioning
of physiological systems and organs of the living organism 相似文献
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