荧光原位杂合(fluorescent in situ hybridization,FISH) 是一种细胞遗传学技术,可以用来对核酸进行检测和定位。荧光标记的核酸探针只和具有高度相似性的核酸杂合,可用于染色体上基因的定位,或在分子生态学中用来标记不同分类细菌或古菌中的核糖体RNA。
在医院的妇产科或相关检验所中,会利用荧光原位杂合技术来判别胎儿的染色体是否正常。
对于利用rRNA的荧光原位杂合来说,如下原因可导致较低的荧光信号强度:
- 较低的细胞核糖体含量
- 较低的细胞周边的通透性
- 较低的目标序列可接触性(由于rRNA的折叠产生的构象,有些位置与rRNA分子内其他链或其他rRNA或蛋白紧密接触,从而使探针无法和目标序列杂合)
为检验细胞中的目标序列是否容易被探针杂合,及测试最佳杂合温度,可利用“克隆荧光原位杂合”(clone-FISH)进行试验:将rRNA基因结合入质粒,转化至大肠杆菌中表达,构成核糖体,再用荧光标记的探针杂合。
FISH可与流式细胞术联用,对特定荧光标记的细胞进行计数或者分离。
变体
- 酶联荧光原位杂合(CARD-FISH)
延伸阅读
- Pernthaler A, Pernthaler J, Amann R. Fluorescence In Situ Hybridization and Catalyzed Reporter Deposition for the Identification of Marine Bacteria. Applied and Environmental Microbiology. 2002, 68 (6): 3094–3101. PMC 123953 . PMID 12039771. doi:10.1128/AEM.68.6.3094-3101.2002.
- Wagner M, Horn M, Daims H. Fluorescence in situ hybridisation for the identification and characterisation of prokaryotes. Current Opinion in Microbiology. 2003, 2003 (6): 302–309. doi:10.1016/S1369-5274(03)00054-7.
- Carthy, J.D. Viewpoints In Biology. England: Butterworth & Co. 1965: 66.
外部链接
- 医学主题词表(MeSH):Fluorescence+in+Situ+Hybridization
- Information on fiber FISH from the Olympus Corporation
- A guide to fiber FISH (页面存档备份,存于互联网档案馆) from Octavian Henegariu
- Fibre FISH protocol (页面存档备份,存于互联网档案馆) from the Human Genome Project at the Sanger Centre
- CARD-FISH, BioMineWiki (页面存档备份,存于互联网档案馆)
- Preparation of Complex DNA Probe Sets for 3D FISH with up to Six Different Fluorochromes (页面存档备份,存于互联网档案馆)
- FISH technical notes and protocols from GeneDetect.com (页面存档备份,存于互联网档案馆)
- Fluorescence in situ Hybridization Photos of bacteria (页面存档备份,存于互联网档案馆)
- Rational design of polynucleotide probe mixes to identify particular genes in defined taxa: www.dnaBaser.com/PolyPro
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