Birkenmeier E H, Schneider U, Thurston S J
Jackson Laboratory, Bar Harbor, Maine 04609.
Mamm Genome. 1992;3(10):537-45. doi: 10.1007/BF00350618.
PCR primers of arbitrary nucleotide sequence have identified DNA polymorphisms useful for genetic mapping in a large variety of organisms. Although technically very powerful, the use of arbitrary primers for genome mapping has the disadvantage of characterizing DNA sequences of unknown function. Thus, there is no reason to anticipate that DNA fragments amplified by use of arbitrary primers will be enriched for either transcribed or promoter sequences that may be conserved in evolution. For these reasons, we modified the arbitrarily primed PCR method by using oligonucleotide primers derived from conserved promoter elements and protein motifs. Twenty-nine of these primers were tested individually and in pairwise combinations for their ability to amplify genomic DNA from a variety of species including various inbred strains of laboratory mice and Mus spretus. Using recombinant inbred strains of mice, we determined the chromosomal location of 27 polymorphic fragments in the mouse genome. The results demonstrated that motif sequence-tagged PCR products are reliable markers for mapping the mouse genome and that motif primers can also be used for genomic fingerprinting of many divergent species.
具有任意核苷酸序列的PCR引物已在多种生物体中鉴定出可用于基因图谱绘制的DNA多态性。尽管在技术上非常强大,但使用任意引物进行基因组图谱绘制的缺点是表征功能未知的DNA序列。因此,没有理由预期使用任意引物扩增的DNA片段会富集可能在进化中保守的转录或启动子序列。出于这些原因,我们通过使用源自保守启动子元件和蛋白质基序的寡核苷酸引物对任意引物PCR方法进行了改进。对其中29种引物进行了单独测试,并进行了两两组合测试,以检测它们从包括实验室小鼠的各种近交系和小家鼠在内的多种物种中扩增基因组DNA的能力。利用小鼠的重组近交系,我们确定了小鼠基因组中27个多态性片段的染色体位置。结果表明,基序序列标签PCR产物是绘制小鼠基因组图谱的可靠标记,并且基序引物也可用于许多不同物种的基因组指纹分析。