Kazianis S, Morizot D C, McEntire B B, Nairn R S, Borowsky R L
Department of Biology, New York University, New York 10003, USA.
Genome Res. 1996 Apr;6(4):280-9. doi: 10.1101/gr.6.4.280.
The combined use of the arbitrarily primed polymerase chain reaction [AP-PCR, also known as random amplification of polymorphic DNA (RAPD)] and isozyme mapping resulted in the production of 87 potential marker loci, enabling an overall expansion within the genetic map of the fish genus Xiphophorus. Use of DNA sequencing-style acrylamide gels and carefully controlled conditions of amplification and silver staining allowed exceptional resolution and reproducibility of AP-PCR/RAPD generated markers. Linkage analysis of AP-PCR/RAPD and isozyme markers resulted in the addition of 16 new markers to Xiphophorus linkage groups (LGs) I, II, III, V, IX, X, XII, and XIV. Addition of 5 AP-PCR/RAPD markers to linkage group U6 containing the Tailspot pigment pattern locus (P) and designation of eight new unassigned linkage groups with 22 markers was also accomplished. Genetic linkage data allowed inference of the existence of a novel pigment pattern modifier locus. Expansion of the Xiphophorus gene map by linkage analysis of AP-PCR/RAPD markers in conjunction with isozyme polymorphisms should lead to the rapid saturation of genetic linkage groups such as LG V, which will probably be instrumental to cloning the Diff tumor suppressor gene locus.
任意引物聚合酶链反应[AP-PCR，也称为随机扩增多态性DNA（RAPD）]与同工酶图谱分析的联合应用，产生了87个潜在的标记位点，从而使剑尾鱼属的遗传图谱得到了全面扩展。使用DNA测序型聚丙烯酰胺凝胶以及严格控制的扩增和银染条件，使得AP-PCR/RAPD产生的标记具有出色的分辨率和可重复性。对AP-PCR/RAPD和同工酶标记进行连锁分析，为剑尾鱼的连锁群（LGs）I、II、III、V、IX、X、XII和XIV增加了16个新标记。还将5个AP-PCR/RAPD标记添加到了包含尾斑色素模式位点（P）的连锁群U6中，并确定了8个新的未分配连锁群，其中有22个标记。遗传连锁数据有助于推断出一个新的色素模式修饰位点的存在。通过对AP-PCR/RAPD标记与同工酶多态性进行连锁分析来扩展剑尾鱼基因图谱，应能使遗传连锁群（如LG V）迅速饱和，这可能有助于克隆Diff肿瘤抑制基因位点。
