Ellegren H, Chowdhary B, Johansson M, Andersson L
Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences, Uppsala.
Anim Genet. 1994 Jun;25(3):155-64. doi: 10.1111/j.1365-2052.1994.tb00104.x.
An essential part in the development of informative linkage maps is to include genetic markers that have been anchored by physical mapping. Here a set of 18 porcine cosmid-derived genetic markers are reported that have been mapped by linkage analysis, and that also have been physically localized by fluorescence in situ hybridization (FISH). Three different strategies were used to establish polymorphic markers from the cosmid clones. Firstly, dinucleotide microsatellite loci were derived by sequencing cosmid subclones containing (CA)n repeats. Secondly, variable SINE 3' poly(A) tracts (SINEVA) were identified by direct SINE-PCR amplification of cosmid clones. Thirdly, the cosmids were used in Southern blot hybridization to detect restriction fragment length polymorphisms (RFLPs). Compared with the most recent consensus compilation of the porcine gene map, the present assignment of markers to chromosomes 2p, 3, 4, 10, 12q, and 16 represents the first loci mapped to these chromosomes, for which linkage as well as in situ data are now available.
构建信息丰富的连锁图谱的一个关键部分是纳入已通过物理图谱定位的遗传标记。本文报道了一组18个猪黏粒衍生的遗传标记,这些标记已通过连锁分析进行定位,并且也已通过荧光原位杂交(FISH)进行了物理定位。采用了三种不同的策略从黏粒克隆中建立多态性标记。首先,通过对含有(CA)n重复序列的黏粒亚克隆进行测序获得二核苷酸微卫星位点。其次,通过对黏粒克隆进行直接SINE-PCR扩增鉴定可变SINE 3'聚腺苷酸序列(SINEVA)。第三,将黏粒用于Southern杂交以检测限制性片段长度多态性(RFLP)。与猪基因图谱的最新共识汇编相比,目前将标记定位到2p、3、4、10、12q和16号染色体上代表了首次定位到这些染色体上的位点,现在已有这些位点的连锁和原位数据。