Amar L C, Arnaud D, Cambrou J, Guenet J L, Avner P R
EMBO J. 1985 Dec 30;4(13B):3695-700. doi: 10.1002/j.1460-2075.1985.tb04137.x.
Two libraries enriched in murine X chromosome material have been constructed in the lambda vector NM 1149 from flow-sorted chromosomes. Inserts of unique genomic sequence DNA were purified and their X chromosome specificity characterised by hybridisation to a panel of somatic cell hybrid lines. Of the first five such X chromosome-specific probes characterised, all detect restriction fragment length polymorphisms (RFLPs) between inbred mouse laboratory strains such as C57BL/6 and BALB/c and the SPE/Pas mouse strain established from a wild Mus spretus mouse, when their DNAs are digested with the restriction enzyme TaqI. Taking advantage of these RFLPs, all five probes have been localised on the X chromosome using an interspecific backcross between the B6CBARI and SPE/Pas mouse strains segregating the X chromosome markers hypoxanthine phosphoribosyl transferase (Hprt) and Tabby (Ta). Three of the probes map to the region between the centromere and Hprt, and two distal to Ta. Since such X-specific sequence probes detect RFLPs between M. spretus and M. musculus domesticus DNAs with high frequency, a large panel of well localised probes should soon be available for studies of biological problems associated with the X chromosome which can best be approached using the murine species.
已从流式细胞分选的染色体中,以λ噬菌体载体NM 1149构建了两个富含小鼠X染色体物质的文库。纯化了独特基因组序列DNA的插入片段,并通过与一组体细胞杂交系杂交来表征其X染色体特异性。在所表征的前五个此类X染色体特异性探针中,当用限制性内切酶TaqI消化C57BL/6和BALB/c等近交小鼠实验室品系以及从野生斯氏小家鼠建立的SPE/Pas小鼠品系的DNA时,所有探针均检测到限制性片段长度多态性(RFLP)。利用这些RFLP,使用在分离X染色体标记次黄嘌呤磷酸核糖基转移酶(Hprt)和虎斑(Ta)的B6CBARI和SPE/Pas小鼠品系之间的种间回交,将所有五个探针定位在X染色体上。其中三个探针定位于着丝粒和Hprt之间的区域,另外两个定位于Ta的远端。由于此类X特异性序列探针能高频检测斯氏小家鼠和小家鼠DNA之间的RFLP,因此很快就会有一大组定位良好的探针可用于研究与X染色体相关的生物学问题,而使用小鼠物种能最好地解决这些问题。