基于RAPD改进的分子标记技术

Modified molecular marking techniques based on RAPD.

随机扩增多态DNA(RAPD)由于经济、快速简便、不需预知模板DNA信息、对模板DNA要求较低,因此被大量应用于昆虫学的研究中,如昆虫的鉴定分类、构建连锁图谱、系统发育分析等。但RAPD也有许多缺点,主要是一种显性标记而不能区分杂合体和纯合体,结果的重复性易受多因素的影响等。针对这些缺点,人们通过将RAPD与其它检测技术如聚丙烯酰胺凝胶电泳、单链构象多态(SSCP)、变性梯度凝胶电泳(DGGE)等相结合,提高了其分析的灵敏度和稳定性;再者,还将RAPD标记发展成为其它更稳定的或共显性的标记,如SCAR、微卫星等,为RAPD进一步的应用开阔了前景。文章对这些检测和转化的改进技术进行综述。

Random amplified polymorphic DNA（RAPD）technique is economical,fast and easy to handle.It requires only small amount of material and no prior information about template DNA.This technique have been widely used for many types of insect genetic analyses,including different taxa differentiation,genome mapping,genotype fingerprinting,phylogeny reconstruction,and measuring genetic polymorphisms.However,RAPD also has some disadvantages,especially the majority of its alleles segregated as dominant markers,which could not differentiate homozygotes and heterozygotes,and its repeatability was sensitive to PCR conditions.In order to overcome these disadvantages,RAPD is combined with other techniques such as polyacrylamide gel electrophoresis,single-strand conformation polymorphism（SSCP）,denaturing gradient gel electrophoresis（DGGE）and so on to enhance its sensitivity and reliability.On the other hand,more reliable or codominant markers based on RAPD are also adopted,such as sequence characterized amplified regions（SCAR）,microsatellite,etc.This paper reviewed current progress in the modifications and improvements on the RAPD technique.