Reed K M, Mendoza K M, Beattie C W
Department of Veterinary Pathobiology, University of Minnesota, St. Paul 55108, USA.
Genome. 2000 Oct;43(5):796-802.
Cross-species amplification of 520 chicken microsatellite markers was tested by polymerase chain reaction with genomic DNA of the turkey (Meleagris gallopavo). Each primer pair was tested at six different combinations of annealing temperature and MgCl2 concentration. A total of 280 (54%) of the primer pairs produced amplification products. The majority of these products were similar, if not identical in size to those expected based on the fragment sizes of the corresponding chicken loci. Structure of the dinucleotide repeat and flanking sequences was examined for 13 turkey fragments (amplified with chicken primers) and 5 chicken fragments (amplified with turkey primers). Sequence analysis found a wide array of mutations between species in addition to differences in repeat length. To estimate the usefulness of the amplified loci for genetic mapping in the turkey, allelic polymorphism was determined for 57 of the 280 amplified loci. A total of 20 of 57 markers (35%) were polymorphic with an average of 1.4 alleles per locus. The results of this study suggest that approximately 20% of the chicken microsatellite markers will be useful for mapping the turkey genome.
利用火鸡(Meleagris gallopavo)的基因组DNA,通过聚合酶链反应对520个鸡微卫星标记进行了种间扩增测试。每个引物对在退火温度和MgCl2浓度的六种不同组合下进行测试。总共280对(54%)引物对产生了扩增产物。这些产物中的大多数,即使大小不完全相同,也与基于相应鸡基因座片段大小预期的产物相似。对13个火鸡片段(用鸡引物扩增)和5个鸡片段(用火鸡引物扩增)的二核苷酸重复序列和侧翼序列结构进行了研究。序列分析发现,除了重复长度的差异外,物种之间还存在大量突变。为了评估扩增基因座在火鸡遗传图谱构建中的实用性,对280个扩增基因座中的57个进行了等位基因多态性测定。57个标记中有20个(35%)具有多态性,每个基因座平均有1.4个等位基因。本研究结果表明,约20%的鸡微卫星标记可用于火鸡基因组图谱的构建。