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植物中 ,程序性细胞死亡 (PCD)发生在植物生殖和发育的许多方面 ,已有的研究表明 ,在玉米种子的发育过程中 ,胚乳组织经历了程序性细胞死亡的过程。bz1 (bronze)和bz2是与种子的糊粉层发育相关的花青素生物合成基因 ,在玉米基因组中 ,bz1基因所在区域是重组热点 ,bz2与类黄酮的酰化、糖基化、转运、沉积等有关 ,基因的物理定位有利于基因的分离和克隆。TSA FISH (Tyramidesignalamplificationfluorescenceinsituhybridization)是一种新颖的高灵敏度的荧光原位杂交技术 ,它的主要反应原理是辣根过氧化物酶催化过氧化氢和标记的酪胺分子 (tyramide)的苯环部分反应 ,使荧光标记的酪胺分子在直接带有或间接带有HRP报告分子的探针周围沉积 ,信号因此得以极大的放大 ,从而大大提高了荧光原位杂交技术的灵敏度 ,90年代中期开始引入动物和人类组织化学和细胞遗传学研究中 ,2 0 0 1年才应用于植物细胞遗传学的研究。利用这一技术 ,我们将bz1基因定位于玉米的第 9染色体的短臂和第 1染色体的长臂上 ,其信号点距着丝粒的百分距离分别为 40 .2 ,75 .4;bz2基因定位于玉米的第 1染色体的长臂和第 5染色体的短臂上 ,其信号点距着丝粒的百分距离分别为2 1 .6,1 5 .3。本文讨论了TSA FISH技术在植物中小的、低拷贝的DNA序
In plants, programmed cell death (PCD) occurs in many aspects of plant reproduction and development. Studies have shown that endosperm tissue undergoes programmed cell death during the development of maize seeds. bz1 (bronze) and bz2 are anthocyanin biosynthetic genes related to aleurone development of seeds. In the maize genome, the region of bz1 gene is a recombination hot spot, acylation, glycosylation, transport of bz2 with flavonoids, Sedimentation and other related gene physical positioning is conducive to gene isolation and cloning. TSA FISH (Tyramides signaling amplification fluorescence in situ hybridization) is a novel high-sensitivity fluorescence in situ hybridization. Its main reaction principle is that horseradish peroxidase catalyzes the partial reaction of hydrogen peroxide with the benzene ring of labeled tyramide, Fluorescently labeled tyramide molecules are deposited around probes directly or indirectly with HRP reporter and the signal is greatly amplified thereby greatly increasing the sensitivity of fluorescence in situ hybridization and the introduction of animals into the mid-1990s And human tissue chemistry and cytogenetics, 2001 was applied to plant cytogenetics. Using this technique, we mapped the bz1 gene to the short arm of chromosome 9 and the long arm of chromosome 1 in maize, and their signal points were separated from the centromere by 40.2, 75.4, respectively. Bz2 The gene was located on the long arm of chromosome 1 and the short arm of chromosome 5 in maize. The distance between the signal points and the centromere was 21.6, 15.5, respectively. This article discusses the small, low-copy DNA sequence of TSA FISH technology in plants