High-resolution physical mapping of human 5q31-q33 using three methods: radiation hybrid mapping, interphase fluorescence in situ hybridization, and pulsed-field gel electrophoresis.
High-resolution physical mapping of human 5q31-q33 using three methods: radiation hybrid mapping, interphase fluorescence in situ hybridization, and pulsed-field gel electrophoresis.
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DOI：
10.1006/geno.1994.1636
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发表时间：
1994-11
期刊：
影响因子：
4.4
通讯作者：
J. Warrington;U. Bengtsson
中科院分区：
文献类型：
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作者：
J. Warrington;U. Bengtsson
用辐射杂交(RH)作图、脉冲场凝胶电泳法(PFGE)和间期核荧光原位杂交(FISH)三种物理作图方法确定了人类5号染色体Q31-Q33区域12个基因座之间的顺序和相对距离，并比较了每种方法所获得的信息是否一致。根据数据组合，预测的12个基因座的顺序为：cen-ADRB2-PDEA-CSF1R-RPS14-ANX6-SPARC+-GLRA1-GLUR1-ADRA1B-IL12-GABRG2-GABRA1-TEL。在这些基因座跨越的5-Mb区域，我们确定一个RH定位单位(centiray，cR6500)相当于21kb，与以前使用相同的辐射杂交来定位其他区域的27-34kb/cR6500的估计很好地一致。在这三种方法中，RH映射是迄今为止确定顺序和距离的最简单和最有效的方法。此外，RH映射的分辨率范围最宽，从大约200kb到几兆空间不等。间期FISH是澄清紧密连锁标记顺序的最有信息量的方法。这项研究得到的FISH距离将作为高分辨率定位人类5号染色体长臂上其他区域的内部参考。与RH作图和FISH相比，PFGE是三种方法中最不可靠的，产生的一些数据与其他两种方法测定的距离不一致。
Three physical mapping methods, radiation hybrid (RH) mapping, pulsed-field gel electrophoresis (PFGE), and fluorescence in situ hybridization (FISH) of interphase nuclei, were used to determine the order and relative distances between 12 loci in the q31-q33 region of human chromosome 5. The information obtained by each of the methods was compared to determine whether they gave consistent results. Based upon a combination of data, the predicted order of the 12 loci is cen-ADRB2-PDEA-CSF1R-RPS14-ANX6-SPARC++ +-GLRA1-GLUR1-ADRA1B-IL12- GABRG2-GABRA1-tel. Over the 5-Mb region spanned by these loci, we determined that an RH mapping unit (centiray, cR6500) is equivalent to 21 kb, in good agreement with previous estimates of 27-34 kb/cR6500 using the same set of radiation hybrids to map other regions. Of the three methods, RH mapping was by far the easiest and most efficient method for determining both order and distances. In addition, the range of resolution of RH mapping was the broadest, ranging from approximately 200 kb to several megabasepairs. Interphase FISH was the most informative method for clarifying the order of closely linked markers. The FISH distances obtained in this study will be useful as an internal reference for high-resolution mapping studies of other regions on the long arm of human chromosome 5. In contrast to RH mapping and FISH, PFGE was the least reliable of the three methods, producing some data that were inconsistent with the distances determined by the other two methods.