Bowden D W, Müller-Kahle H, Gravius T C, Helms C, Watt-Morgan D, Green P, Donis-Keller H
Human Genetics Department, Collaborative Research, Incorporated, Waltham, MA 02154.
Am J Hum Genet. 1989 May;44(5):671-8.
As part of our search for polymorphic DNA probes, we have screened cosmids from a human genomic DNA library for their ability to reveal RFLPs. A total of 101 randomly isolated cosmid clones were tested in Southern hybridizations for polymorphic band patterns. Fifty-four of these clones revealed RFLPs with one or more of nine restriction enzymes. Twenty-three of these clones have been further characterized and assigned to 10 different chromosomes by linkage analysis or by hybridization to panels of human-hamster hybrid cell lines. Fifteen of the probes have heterozygosities greater than or equal to .5. The relative efficiency of RsaI and PstI restriction enzymes in detecting polymorphism was different from results obtained with libraries constructed in bacteriophage vectors. Screening randomly selected cosmid probes is an efficient method for detecting RFLPs.
作为寻找多态性DNA探针工作的一部分,我们从一个人类基因组DNA文库中筛选黏粒,检测其揭示限制性片段长度多态性(RFLP)的能力。总共101个随机分离的黏粒克隆在Southern杂交中检测多态性条带模式。其中54个克隆与9种限制性内切酶中的一种或多种显示出RFLP。这些克隆中有23个已通过连锁分析或与人-仓鼠杂交细胞系面板杂交作了进一步鉴定,并定位到10条不同的染色体上。15个探针的杂合度大于或等于0.5。RsaI和PstI限制性内切酶在检测多态性方面的相对效率与用噬菌体载体构建的文库所得到的结果不同。筛选随机选择的黏粒探针是检测RFLP的一种有效方法。