用生物素（Biotin-16-dUTP)标记的大麦Betzes基因组DNA作探针，以普通小麦中国春总DNA作封阻进行基因组原位杂交（Genome in situ hybridization,简称GISH），从13株小麦-大麦杂交后代中鉴定出2个含有3条大麦Betzes 2H染色体的材料（2n...用生物素（Biotin-16-dUTP)标记的大麦Betzes基因组DNA作探针，以普通小麦中国春总DNA作封阻进行基因组原位杂交（Genome in situ hybridization,简称GISH），从13株小麦-大麦杂交后代中鉴定出2个含有3条大麦Betzes 2H染色体的材料（2n＝43）；2个2H单体异代换系（2n＝42）；7个2H二体异代换系（2n＝42）。用已定位在小麦第2部分同源群短臂上的探针psr131进行RFLP分析，结果表明大麦Betzes、代换系A5有1条区别于小麦中国春的特异带，A5的2条2A染色体被大麦Betzes的2条2H染色体所代换。GISH及RFLP的准确鉴定及小麦－大麦2H二体异代换系的首次获得，为向小麦导入2H染色体上的有用基因提供了宝贵材料。展开更多
The uptodate advances of genomic in situ hybridization (GISH) for last ten years are reviewed in the article. These methods initially deriving from animal research have been developed in plant study, such as Multicolo...The uptodate advances of genomic in situ hybridization (GISH) for last ten years are reviewed in the article. These methods initially deriving from animal research have been developed in plant study, such as Multicolor Fluorescence in situ Hybridization (McFISH), Third strand in situ Hybridization (TISH), Comparative Genomic Hybridization (CGH) and Simultaneous in situ Hybridization. GISH applied widely in plant can reveal the species origin and evolution and determine the truth of distant hybrid containing foreign chromatin. Also it can research the chromosome behaviour including B chromosome and explore the genomic mapping and genic function. Finally the applicability and resolving power of GISH about taxonomic distance, genomic size and the length of chromosome fragment are discussed, as well as the future and the status of GISH in plant.展开更多
在基因组原位杂交中,适当的封阻可以大大提高基因组原位杂交的效率。本研究采用煮沸法、超声波剪切法对大白菜基因组DNA进行剪切,研究了大白菜封阻DNA的制备方法。结果表明:珠沸法效率高,操作简单,当煮沸70 min DNA片段大小主要集中在20...在基因组原位杂交中,适当的封阻可以大大提高基因组原位杂交的效率。本研究采用煮沸法、超声波剪切法对大白菜基因组DNA进行剪切,研究了大白菜封阻DNA的制备方法。结果表明:珠沸法效率高,操作简单,当煮沸70 min DNA片段大小主要集中在200～500 bp,适于在基因组原位杂交中作为封阻。研究结果为基因组原位杂交的应用奠定了基础。展开更多
采用生物素标记的拟南芥基因组DNA探针在75%杂交严谨度下对双子叶植物番茄、蚕豆和单子叶植物水稻、玉米、大麦的染色体进行了比较基因组荧光原位杂交(comparative genomic in situ hybridization,cGISH)分析,以揭示拟南芥与远缘植物基...采用生物素标记的拟南芥基因组DNA探针在75%杂交严谨度下对双子叶植物番茄、蚕豆和单子叶植物水稻、玉米、大麦的染色体进行了比较基因组荧光原位杂交(comparative genomic in situ hybridization,cGISH)分析,以揭示拟南芥与远缘植物基因组间的同源性.cGISH信号代表了拟南芥基因组DNA中的重复DNA与靶物种染色体上同源序列的杂交.探针DNA在所有靶物种的全部染色体上都产生了杂交信号.杂交信号为散在分布,并呈现随基因组增大,杂交信号增多,且分布更加分散的趋势.所有靶物种的核仁组织区(NOR)都显示了明显强于其他区域的杂交信号,表明拟南芥基因组DNA探针可用于植物NOR的物理定位.在所有的靶物种中,信号主要分布在染色体的臂中间区和末端,着丝粒或近着丝粒区有少数信号分布.大麦染色体显示了与C-和N-带不同的独特的cGISH信号带型,表明此探针可用于不同植物染色体的识别.这些结果表明,拟南芥基因组与远缘植物基因组之间,除rDNA和端粒重复序列外,还存在其它同源的重复DNA;一些重复DNA序列在被子植物分歧进化为单子叶和双子叶植物之前就已存在,虽经历了长期的进化过程,至今在远缘物种之间仍保持了较高的同源性.结果还提示,大基因组中古老而保守的重复DNA在进化过程中发生了明显的扩增.展开更多
文摘用生物素（Biotin-16-dUTP)标记的大麦Betzes基因组DNA作探针，以普通小麦中国春总DNA作封阻进行基因组原位杂交（Genome in situ hybridization,简称GISH），从13株小麦-大麦杂交后代中鉴定出2个含有3条大麦Betzes 2H染色体的材料（2n＝43）；2个2H单体异代换系（2n＝42）；7个2H二体异代换系（2n＝42）。用已定位在小麦第2部分同源群短臂上的探针psr131进行RFLP分析，结果表明大麦Betzes、代换系A5有1条区别于小麦中国春的特异带，A5的2条2A染色体被大麦Betzes的2条2H染色体所代换。GISH及RFLP的准确鉴定及小麦－大麦2H二体异代换系的首次获得，为向小麦导入2H染色体上的有用基因提供了宝贵材料。
文摘The uptodate advances of genomic in situ hybridization (GISH) for last ten years are reviewed in the article. These methods initially deriving from animal research have been developed in plant study, such as Multicolor Fluorescence in situ Hybridization (McFISH), Third strand in situ Hybridization (TISH), Comparative Genomic Hybridization (CGH) and Simultaneous in situ Hybridization. GISH applied widely in plant can reveal the species origin and evolution and determine the truth of distant hybrid containing foreign chromatin. Also it can research the chromosome behaviour including B chromosome and explore the genomic mapping and genic function. Finally the applicability and resolving power of GISH about taxonomic distance, genomic size and the length of chromosome fragment are discussed, as well as the future and the status of GISH in plant.
文摘在基因组原位杂交中,适当的封阻可以大大提高基因组原位杂交的效率。本研究采用煮沸法、超声波剪切法对大白菜基因组DNA进行剪切,研究了大白菜封阻DNA的制备方法。结果表明:珠沸法效率高,操作简单,当煮沸70 min DNA片段大小主要集中在200～500 bp,适于在基因组原位杂交中作为封阻。研究结果为基因组原位杂交的应用奠定了基础。
文摘采用生物素标记的拟南芥基因组DNA探针在75%杂交严谨度下对双子叶植物番茄、蚕豆和单子叶植物水稻、玉米、大麦的染色体进行了比较基因组荧光原位杂交(comparative genomic in situ hybridization,cGISH)分析,以揭示拟南芥与远缘植物基因组间的同源性.cGISH信号代表了拟南芥基因组DNA中的重复DNA与靶物种染色体上同源序列的杂交.探针DNA在所有靶物种的全部染色体上都产生了杂交信号.杂交信号为散在分布,并呈现随基因组增大,杂交信号增多,且分布更加分散的趋势.所有靶物种的核仁组织区(NOR)都显示了明显强于其他区域的杂交信号,表明拟南芥基因组DNA探针可用于植物NOR的物理定位.在所有的靶物种中,信号主要分布在染色体的臂中间区和末端,着丝粒或近着丝粒区有少数信号分布.大麦染色体显示了与C-和N-带不同的独特的cGISH信号带型,表明此探针可用于不同植物染色体的识别.这些结果表明,拟南芥基因组与远缘植物基因组之间,除rDNA和端粒重复序列外,还存在其它同源的重复DNA;一些重复DNA序列在被子植物分歧进化为单子叶和双子叶植物之前就已存在,虽经历了长期的进化过程,至今在远缘物种之间仍保持了较高的同源性.结果还提示,大基因组中古老而保守的重复DNA在进化过程中发生了明显的扩增.
