Toldo S S, Fries R, Steffen P, Neibergs H L, Barendse W, Womack J E, Hetzel D J, Stranzinger G
Department of Animal Science, Swiss Federal Institute of Technology, ETH Zentrum, Zurich.
Mamm Genome. 1993 Dec;4(12):720-7. doi: 10.1007/BF00357796.
To identify physical and genetic anchor loci on bovine chromosomes, 13 cosmids, obtained after the screening of partial bovine cosmid libraries with the (CA)n microsatellite motif, were mapped by fluorescence in situ hybridization (FISH). Eleven cosmid probes yielded a specific signal on one of the bovine chromosomes and identified the following loci: D5S2, D5S3, D6S3, D8S1, D11S5, D13S1, D16S5, D17S2, D19S2, D19S3, D21S8. Two cosmids produced centromeric signals on many chromosomes. The microsatellite-containing regions were subcloned and sequenced. The sequence information revealed that the two centromeric cosmids were derived from bovine satellites 1.723 and 1.709, respectively. A cosmid located in the subtelomeric region of Chromosome (Chr) 17 (D17S2) had features of a chromosome-specific satellite. Primers were designed for eight of the nonsatellite cosmids, and seven of these microsatellites were polymorphic with between three and eight alleles on a set of outbred reference families. The polymorphic and chromosomally mapped loci can now be used to physically anchor other bovine polymorphic markers by linkage analysis. The microsatellite primers were also applied to DNA samples of a previously characterized panel of somatic hybrid cell lines, allowing the assignment of seven microsatellite loci to defined syntenic groups. These assignments confirmed earlier mapping results, revealed a probable case of false synteny, and placed two formerly unassigned syntenic groups on specific chromosomes.
为了确定牛染色体上的物理和遗传锚定基因座,用(CA)n微卫星基序筛选部分牛黏粒文库后获得的13个黏粒,通过荧光原位杂交(FISH)进行定位。11个黏粒探针在一条牛染色体上产生了特异性信号,并鉴定出以下基因座:D5S2、D5S3、D6S3、D8S1、D11S5、D13S1、D16S5、D17S2、D19S2、D19S3、D21S8。两个黏粒在许多染色体上产生着丝粒信号。对含微卫星的区域进行亚克隆并测序。序列信息显示,这两个着丝粒黏粒分别来源于牛卫星1.723和1.709。位于17号染色体(Chr)亚端粒区域的一个黏粒(D17S2)具有染色体特异性卫星的特征。为8个非卫星黏粒设计了引物,其中7个微卫星在一组远交参考家系中具有3至8个等位基因的多态性。现在可以通过连锁分析,利用这些多态性且已进行染色体定位的基因座,对其他牛多态性标记进行物理锚定。微卫星引物还应用于一组先前已鉴定的体细胞杂种细胞系的DNA样本,从而将7个微卫星基因座分配到特定的同线群中。这些分配结果证实了早期的定位结果,揭示了一个可能的假同线性案例,并将两个以前未分配的同线群定位到特定染色体上。