Romans P, Seeley D C, Kew Y, Gwadz R W
Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
J Med Entomol. 1991 Jan;28(1):147-51. doi: 10.1093/jmedent/28.1.147.
Analysis of restriction fragment length polymorphism (RFLP) is a powerful tool for analyzing linkage relationships in species where few genetic markers have been described and where conduct of crosses is difficult. It also permits integration of genetic and physical (cytogenetic) data when the probes have been mapped by in situ hybridization. To illustrate the utility of the method, and because some mutations of a diphenol oxidase gene might conceivably produce the malaria refractoriness phenotype of ookinete-oocyst encapsulation, backcrosses between two inbred lines of Anopheles gambiae Giles were carried out to determine the linkage relationship between the diphenol oxidase A2 (Dox) gene and the esterase locus associated with refractoriness to Plasmodium cynomolgi NIH. The Dox alleles were a Sal I restriction fragment length polymorphism visualized by probing Southern blotted DNA from portions of individual mosquitoes with a cloned Dox gene probe. The two genes were shown to segregate independently.
限制性片段长度多态性（RFLP）分析是一种强大的工具，可用于分析那些遗传标记描述较少且杂交操作困难的物种中的连锁关系。当通过原位杂交对探针进行定位时，它还能实现遗传数据与物理（细胞遗传学）数据的整合。为了说明该方法的实用性，并且由于双酚氧化酶基因的某些突变可能会产生疟原虫卵囊包囊化的疟疾抗性表型，我们对冈比亚按蚊（Anopheles gambiae Giles）的两个近交系进行了回交，以确定双酚氧化酶A2（Dox）基因与与对食蟹猴疟原虫NIH抗性相关的酯酶基因座之间的连锁关系。Dox等位基因是一种Sal I限制性片段长度多态性，通过用克隆的Dox基因探针探测单个蚊子部分的Southern印迹DNA来可视化。结果表明这两个基因是独立分离的。
