共查询到18条相似文献,搜索用时 187 毫秒
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cDNA-AFLP技术及其在植物基因表达研究中的应用 总被引:1,自引:0,他引:1
cDNA-AFLP技术是一种新的研究基因表达的技术,具有重复性好、稳定、可靠的特点,可对生物体转录组进行全面、系统的分析,广泛应用于基因表达特性研究、植物遗传标记分析和分离植物基因等方面.近年来随着技术的不断进步,设备的不断改进,许多新的研究方法不断的产生,该技术取得了迅速的发展.本文就cDNA-AFLP技术的原理、技术特点及其在植物基因表达研究中的应用进行了综述. 相似文献
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本文在简要介绍原位杂交技术的基础之上,重点介绍了该技术在植物遗传育种领域,即在(1)异源染色质及染色体畸变检测;(2)植物基因工程及基因表达研究;(3)构建植物基因物理图谱;(4)染色体RNA研究等方面的应用现状,并对原位杂交技术在提高检出率,与染色体显带技术结合,PCR-原位杂交等方面提了一些见解。 相似文献
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综述了迄今业已分离和鉴定的低温诱导表达的抗冻基因,以及通过染色体基因制图方法揭示和证实的抗冻性基因。并着重地介绍了这些抗冻基因在提高植物抗冻能力上的功能。这些抗冻基因表达合成的新多肽具有高度的亲水性,起着保护和稳定细胞膜结构的作用,从而防止冰冻伤害、提高植物的抗冻能力。同时还介绍了抗冻基因表达中的某些调节因子,其中尤其值得注意的是,CBF基因似乎有可能作为抗冻基因表达的“主开关”,以及Ca2+作为植物细胞的第二信使在传递低温信号、启动和调节抗冻基因表达中可能的重要作用。这些研究结果,不仅为阐明植物的抗冻机理提供了新的证据,而且为改良农作物的抗寒性提供了新的启示。 相似文献
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基因表达调控中的核因子作用 总被引:7,自引:0,他引:7
利用病毒和动物系统对基因表达调控进行了广泛和深入的研究,发现了顺式作用调节序列,鉴定了序列专一的DNA结合蛋白,DNA与蛋白质相互识别、结合及蛋白质与蛋白质相互作用中起作用的蛋白质结构域,并且对调节蛋白基因的克隆和序列进行了分析.基因表达调控领域又由于植物基因调控机制取得的发展而得到了补充,文章着重介绍植物基因中的DNA与蛋白质间的作用;植物调节蛋白基因的分离;这一领域的今后研究方向及展望. 相似文献
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Pudota B. Bhaskar Muthusubramanian Venkateshwaran Lei Wu Jean-Michel Ané Jiming Jiang 《PloS one》2009,4(6)
Potato is the third most important food crop worldwide. However, genetic and genomic research of potato has lagged behind other major crops due to the autopolyploidy and highly heterozygous nature associated with the potato genome. Reliable and technically undemanding techniques are not available for functional gene assays in potato. Here we report the development of a transient gene expression and silencing system in potato. Gene expression or RNAi-based gene silencing constructs were delivered into potato leaf cells using Agrobacterium-mediated infiltration. Agroinfiltration of various gene constructs consistently resulted in potato cell transformation and spread of the transgenic cells around infiltration zones. The efficiency of agroinfiltration was affected by potato genotypes, concentration of Agrobacterium, and plant growth conditions. We demonstrated that the agroinfiltration-based transient gene expression can be used to detect potato proteins in sub-cellular compartments in living cells. We established a double agroinfiltration procedure that allows to test whether a specific gene is associated with potato late blight resistance pathway mediated by the resistance gene RB. This procedure provides a powerful approach for high throughput functional assay for a large number of candidate genes in potato late blight resistance. 相似文献
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Denecke J Aniento F Frigerio L Hawes C Hwang I Mathur J Neuhaus JM Robinson DG 《The Plant cell》2012,24(4):1316-1326
Transient gene expression, in plant protoplasts or specific plant tissues, is a key technique in plant molecular cell biology, aimed at exploring gene products and their modifications to examine functional subdomains, their interactions with other biomolecules, and their subcellular localization. Here, we highlight some of the major advantages and potential pitfalls of the most commonly used transient gene expression models and illustrate how ectopic expression and the use of dominant mutants can provide insights into protein function. 相似文献
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Wheat scab, caused by the fungal pathogen Fusarium graminearum is a devastating disease worldwide. Despite an extensive and coordinated effort to investigate this pathosystem, little progress has been made to understand the molecular basis of host–pathogen interactions, for example how the pathogen causes disease in plant. Recently, a secreted lipase (FGL1) has been identified from the fungus and shown to be an important virulence factor; however, the intrinsic function of FGL1 in plant is unknown. Here, we report the identification of the molecular components that may possibly be involved in the FGL virulence pathway using yeast two hybrid system. FGL gene was amplified from a local virulent strain (F15) and shown to be 99.5% identical to the original published FGL at the amino acid level. We showed that transient expression of this FGL gene by Agroinfiltration in tobacco leaves causes cell death further implicating the role of FGL in virulence. To identify FGL initial physical target in plant, we screened two wheat cDNA libraries using the FGL protein as the bait. From both libraries, a small FKBP-type immunophilin protein, designated wFKBP12, was found to physically interact with FGL. The direct interaction of FGL with wFKBP12 was confirmed in living onion epidermal cells by biomolecular fluorescence complementation (BiFC) assay. To investigate further, we then used wFKBP12 protein as bait and identified an elicitor-responsive protein that contains a potential Ca2 + binding domain. Semi-quantitative PCR showed that this elicitor-responsive gene is down-regulated during the F. graminearum infection suggesting that this protein may be an important component in FGL virulence pathway. This work serves as an initial step to reveal how fungal lipases act as a general virulence factor. 相似文献
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Stéphanie Robert Moustafa Khalf Marie-Claire Goulet Marc-André D’Aoust Frank Sainsbury Dominique Michaud 《PloS one》2013,8(7)
The expression of clinically useful proteins in plants has been bolstered by the development of high-yielding systems for transient protein expression using agroinfiltration. There is a need now to know more about how host plant development and metabolism influence the quantity and quality of recombinant proteins. Endogenous proteolysis is a key determinant of the stability and yield of recombinant proteins in plants. Here we characterised cysteine (C1A) and aspartate (A1) protease profiles in leaves of the widely used expression host Nicotiana benthamiana, in relation with the production of a murine IgG, C5-1, targeted to the cell secretory pathway. Agroinfiltration significantly altered the distribution of C1A and A1 proteases along the leaf age gradient, with a correlation between leaf age and the level of proteolysis in whole-cell and apoplast protein extracts. The co-expression of tomato cystatin SlCYS8, an inhibitor of C1A proteases, alongside C5-1 increased antibody yield by nearly 40% after the usual 6-days incubation period, up to ∼3 mg per plant. No positive effect of SlCYS8 was observed in oldest leaves, in line with an increased level of C1A protease activity and a very low expression rate of the inhibitor. By contrast, C5-1 yield was greater by an additional 40% following 8- to 10-days incubations in younger leaves, where high SlCYS8 expression was maintained. These findings confirm that the co-expression of recombinant protease inhibitors is a promising strategy for increasing recombinant protein yields in plants, but that further opportunity exists to improve this approach by addressing the influence of leaf age and proteases of other classes. 相似文献
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MUHAMMAD MUBIN IMRAN AMIN LUQMAN AMRAO ROB W. BRIDDON SHAHID MANSOOR 《Molecular Plant Pathology》2010,11(2):245-254
A functional analysis of the V2 protein of two monopartite begomoviruses, Papaya leaf curl virus (PaLCuV) and Cotton leaf curl Kokhran virus (CLCuKoV), has been performed. Expression of the V2 gene from a Potato virus X (PVX) vector resulted in severe leaf curling followed by a hypersensitive response (HR) in Nicotiana benthamiana and N. tabacum , demonstrating that the V2 protein is a pathogenicity determinant and a target of host defence responses. Agroinfiltration of a PVX vector expressing the V2 protein resulted in cell death in the infiltrated area. Subsequently, a systemic HR developed that was associated with the long-distance spread of the virus and led to the death of the plant. V2 amino acid sequences encompassing a conserved putative protein kinase C (PKC) phosphorylation motif were shown to be essential for the elicitation of cell death. In co-inoculation experiments, the transient expression of the C2 protein of PaLCuV or Cotton leaf curl Multan virus under the control of the Cauliflower mosaic virus 35S promoter inhibited the HR induced by V2 in the agroinfiltrated area. These findings demonstrate that the V2 protein of monopartite begomoviruses is a pathogenicity determinant and induces an HR that can be suppressed by the C2 protein. The induction and suppression of HR have been demonstrated previously in bipartite begomoviruses and our results extend this to monopartite begomoviruses. 相似文献