Grosse W M, Kappes S M, McGraw R A
Department of Physiology and Pharmacology, College of Veterinary Medicine, University of Georgia, Athens 30602, USA.
Anim Genet. 2000 Jun;31(3):171-7. doi: 10.1046/j.1365-2052.2000.00625.x.
Bovine expressed sequence tags (ESTs) containing microsatellites are suitable markers for both linkage and comparative maps. We isolated clones from a bovine fetal thigh skeletal muscle cDNA library that were positive for a (CA)10 probe. Thirty individual clones were isolated and characterised by sequencing. Sequences from the 5' and 3' ends of a clone were considered as separate ESTs until a contiguous sequence was identified. A total of 47 ESTs were sequenced from the 5' and/or 3' ends and full sequence was obtained for the 30 clones. BLAST nucleotide analysis identified significant homology to known mammalian coding regions for 31 of the bovine ESTs, 30 of which also matched human ESTs or sequence-tagged sites (STS). The remaining 16 bovine ESTs represented novel transcripts. Microsatellites were isolated in 27 of the ESTs, 11 of which were developed into markers and placed on the MARC bovine linkage map. Human cytogenetic map positions were available for 20 of the 30 human EST orthologs, and a putative bovine map position for 17 of the sequences could be inferred using comparative mapping data. These results demonstrated that mapping bovine ESTs containing microsatellites is a plausible strategy to increase the density of gene markers on the bovine linkage and comparative maps.
含有微卫星的牛表达序列标签(ESTs)是用于连锁图谱和比较图谱的合适标记。我们从一个牛胎儿大腿骨骼肌cDNA文库中分离出对(CA)10探针呈阳性的克隆。分离出30个单个克隆并通过测序进行表征。在确定连续序列之前,一个克隆5'和3'端的序列被视为单独的ESTs。从5'和/或3'端总共测序了47个ESTs,并获得了30个克隆的完整序列。BLAST核苷酸分析确定,31个牛ESTs与已知哺乳动物编码区具有显著同源性,其中30个也与人类ESTs或序列标签位点(STS)匹配。其余16个牛ESTs代表新的转录本。在27个ESTs中分离出微卫星,其中11个被开发成标记并定位到MARC牛连锁图谱上。30个人类EST直系同源物中的20个有人类细胞遗传学图谱位置,利用比较图谱数据可以推断出17个序列的假定牛图谱位置。这些结果表明,绘制含有微卫星的牛ESTs图谱是增加牛连锁图谱和比较图谱上基因标记密度的一种可行策略。