Lyons L A, Laughlin T F, Copeland N G, Jenkins N A, Womack J E, O'Brien S J
Laboratory of Genomic Diversity, National Cancer Institute, Frederick Cancer Research and Development Center, Maryland 21702-1201, USA.
Nat Genet. 1997 Jan;15(1):47-56. doi: 10.1038/ng0197-47.
Precise comparisons of mammalian gene maps require common anchor loci as landmarks for conserved chromosomal segments. Using a computer script that automates DNA sequence database alignments, we designed 410 evolutionarily conserved primer pair sequences which are specific for anchor locus gene amplification from any mammalian species' DNA. Primer pairs were designed to span introns for polymorphism ascertainment, and to include sufficient exonic sequence (25-400 bp) to allow for gene identification. A total of 318 primer pairs were optimized for domestic cats, and 86% of the sequenced feline PCR products showed homology to the gene of primer origin. A screen of 20 mammals from 11 orders revealed that 35-52% of the 318 primers yielded a single PCR product without further optimization suggesting that nearly 75% can be optimized for any eutherian mammal.
哺乳动物基因图谱的精确比较需要共同的锚定基因座作为保守染色体片段的标记。我们使用一个能自动进行DNA序列数据库比对的计算机脚本，设计了410个进化上保守的引物对序列，这些序列可特异性地用于从任何哺乳动物物种的DNA中扩增锚定基因座基因。引物对的设计是跨越内含子以确定多态性，并包含足够的外显子序列（25 - 400 bp）以便进行基因鉴定。总共为家猫优化了318个引物对，86%的测序猫科动物PCR产物与引物来源基因具有同源性。对来自11个目20种哺乳动物的筛选显示，318个引物中有35 - 52%在无需进一步优化的情况下产生单一PCR产物，这表明近75%的引物可针对任何真兽类哺乳动物进行优化。
