Serikawa T, Montagutelli X, Simon-Chazottes D, Guénet J L
Unité de Génétique des Mammifères de l'Institut Pasteur, Paris, France.
Mamm Genome. 1992;3(2):65-72. doi: 10.1007/BF00431248.
Ten single, arbitrarily designed oligodeoxynucleotide primers, with 50-70% (G+C) content, were used to amplify by polymerase chain reaction (PCR) sequences with DNA templates from several mouse species (Mus spretus, Mus musculus musculus, and Mus musculus domesticus), as well as DNA from the laboratory rat (Rattus norvegicus). Eight of these ten primers, used either individually or associated in pairs, generated a total of 13 polymorphic products which were used as genetic markers. All of these polymorphic sequences but one were mapped to a particular mouse chromosome, by use of DNA panels prepared either from interspecific backcross progeny of the type (C57BL/6 x Mus spretus)F1 x C57BL/6 or DNA samples prepared from two sets of recombinant inbred (RI) strains (AKXL and BXD). Six rat-specific DNA segments were also assigned to a particular chromosome with DNA panels prepared from 18 rat/mouse somatic cell hybrids segregating rat chromosomes. From these experiments we conclude that, under precisely standardized PCR conditions, the DNA molecules amplified with these arbitrarily designed primers are useful and reliable markers for genetic mapping in both mouse and rat.
使用了10条单链、任意设计的寡脱氧核苷酸引物,其(G+C)含量为50%-70%,通过聚合酶链反应(PCR)以几种小鼠物种(西班牙小鼠、小家鼠指名亚种和小家鼠)的DNA模板以及实验室大鼠(褐家鼠)的DNA来扩增序列。这10条引物中的8条,单独使用或成对组合使用,共产生了13个多态性产物,用作遗传标记。除了一个之外,所有这些多态性序列都通过使用由(C57BL/6×西班牙小鼠)F1×C57BL/6类型的种间回交后代制备的DNA板或由两组重组近交(RI)品系(AKXL和BXD)制备的DNA样品,定位到了特定的小鼠染色体上。六个大鼠特异性DNA片段也通过使用由18个分离大鼠染色体的大鼠/小鼠体细胞杂种制备的DNA板,定位到了特定的染色体上。从这些实验中我们得出结论,在精确标准化的PCR条件下,用这些任意设计的引物扩增的DNA分子是小鼠和大鼠遗传图谱绘制中有用且可靠的标记。