摘要
通过细胞分裂的同步化、低渗、酶消化、瞬间固定和涂片等制片技术,快速制备水稻根尖体细胞染色体标本.该方法制备出的染色体标本,杂质少、染色体分散程度好,满足了染色体微切割用片的要求.染色体微切割和PCR扩增技术,获得了水稻第4 号染色体DNA 文库.经FISH法的初步鉴定,所获得的DNA 确来自水稻细胞核.这为植物特定染色体(或相应区段)的基因克隆提供方便.
By the treatments of synchronization of meristematic cell division in root tips,hypotonic method,transient fixation, enzyme digestion and smearing technique, well spreaded and low contaminated rice chromosomes were quickly prepared and fitted for chromosome microdissection. No.4 chromosomal DNA library was constructed by means of chromosome microdissection and PCR technique. It was primarily confirmed by FISH technique that the DNA library was really from rice cell nuclei. It will be helpful for cloning plant genes located in specific chromosome or related region.
出处
《复旦学报(自然科学版)》
CAS
CSCD
北大核心
1999年第4期444-446,450,共4页
Journal of Fudan University:Natural Science
基金
国家自然科学基金
复旦大学青年自然科学基金
关键词
水稻
染色体
制片
微切割
PCR扩增
rice chromosome
quick preparation of chromosome slide
microdissection
primer linker
PCR amplification