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Advances in plant gene-targeted and functional markers: a review

机译:植物基因靶向和功能标记的研究进展

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摘要

Public genomic databases have provided new directions for molecular marker development and initiated a shift in the types of PCR-based techniques commonly used in plant science. Alongside commonly used arbitrarily amplified DNA markers, other methods have been developed. Targeted fingerprinting marker techniques are based on the well-established practices of arbitrarily amplified DNA methods, but employ novel methodological innovations such as the incorporation of gene or promoter elements in the primers. These markers provide good reproducibility and increased resolution by the concurrent incidence of dominant and co-dominant bands. Despite their promising features, these semi-random markers suffer from possible problems of collision and non-homology analogous to those found with randomly generated fingerprints. Transposable elements, present in abundance in plant genomes, may also be used to generate fingerprints. These markers provide increased genomic coverage by utilizing specific targeted sites and produce bands that mostly seem to be homologous. The biggest drawback with most of these techniques is that prior genomic information about retrotransposons is needed for primer design, prohibiting universal applications. Another class of recently developed methods exploits length polymorphism present in arrays of multi-copy gene families such as cytochrome P450 and β-tubulin genes to provide cross-species amplification and transferability. A specific class of marker makes use of common features of plant resistance genes to generate bands linked to a given phenotype, or to reveal genetic diversity. Conserved DNA-based strategies have limited genome coverage and may fail to reveal genetic diversity, while resistance genes may be under specific evolutionary selection. Markers may also be generated from functional and/or transcribed regions of the genome using different gene-targeting approaches coupled with the use of RNA information. Such techniques have the potential to generate phenotypically linked functional markers, especially when fingerprints are generated from the transcribed or expressed region of the genome. It is to be expected that these recently developed techniques will generate larger datasets, but their shortcomings should also be acknowledged and carefully investigated.
机译:公共基因组数据库为分子标记的开发提供了新的方向,并引发了植物科学中常用的基于PCR的技术类型的转变。除了常用的任意扩增的DNA标记外,还开发了其他方法。靶向指纹标记技术基于任意扩增的DNA方法的公认实践,但采用了新颖的方法学创新,例如将基因或启动子元件掺入引物中。这些标记通过显性和共显性谱带的同时发生提供良好的可重复性和更高的分辨率。尽管它们具有令人鼓舞的特征,但这些半随机标记物可能会发生碰撞和非同源性问题,类似于随机生成的指纹所发现的问题。存在于植物基因组中的转座因子也可用于产生指纹。这些标记物通过利用特定的靶向位点提供了更高的基因组覆盖率,并产生了大多数似乎同源的条带。这些技术中的大多数的最大缺点是引物设计需要先验的有关反转录转座子的基因组信息,从而禁止普遍应用。另一类最近开发的方法利用了多拷贝基因家族(如细胞色素P450和β-微管蛋白基因)阵列中存在的长度多态性,以提供跨物种的扩增和转移能力。一类特定的标记物利用植物抗性基因的共同特征来产生与给定表型相关的条带,或揭示遗传多样性。基于保守的DNA的策略具有有限的基因组覆盖范围,可能无法揭示遗传多样性,而抗性基因可能处于特定的进化选择之下。还可使用不同的基因靶向方法以及RNA信息的使用,从基因组的功能区和/或转录区产生标记。这样的技术具有产生表型连接的功能标记物的潜力,特别是当从基因组的转录或表达区域产生指纹时。可以预期,这些最近开发的技术将生成更大的数据集,但也应认识到并仔细研究其缺点。

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