A human minisatellite probe reveals RFLPs among individuals of two angiosperms.
A human minisatellite probe reveals RFLPs among individuals of two angiosperms.
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人类小卫星探针揭示了两种被子植物个体之间的 RFLP。
DOI：
10.1093/nar/16.23.11378
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发表时间：
1988
影响因子：
14.9
通讯作者：
Schaal,BA
中科院分区：
文献类型：
--
作者：
Rogstad,SH;Patton,JC;Schaal,BA
一些”DNA指纹”探针能够在与来自选定的四足动物的内切核酸酶消化的DNA的印迹杂交时同时揭示几个中度至高变的基因座（1，2，3）。33.15和33.6探针（1，2）与水稻（4）中的序列杂交;杂交模式的差异仅在品种之间发现，但在品种内的个体之间没有发现。在这里，我们展示了探针33.15（由A.杰弗里斯; 1）揭示了来自被子植物美洲黑杨（Povulus deltoides）（棉白杨;图1泳道1-6）和美洲黑杨（P. tremulodies）（白杨;泳道7、8）的天然群体的个体之间的片段差异。在图1中，唯一的数字指的是不同的个体。标记有数字及其撇号的泳道代表来自同一个体的两个不同分支，并且这种泳道的相似性意味着片段长度是体细胞稳定的。虽然这些结果是初步的，但它们表明探针33.15可以与双子叶植物物种个体之间可变的基因组序列杂交，并且与四足动物（1，2，3）和单子叶植物（4）的结果杂交，表明特定的小卫星家族（1，2）可能在真核生物中广泛存在。研究方法：用HaeIII消化从叶中提取的10 μ g DNA/个体（5），在0.9%agofse凝胶中电泳，并印迹到Nylon MSI上。探针33.15（1，2）用P标记（6）。杂交在1.4 XSSC，0.5%脱脂奶粉溶液（7）中于65 ℃杂交12小时;滤膜在IX Denhardts，2 XSSC，0.2%SDS中洗涤（两次15分钟洗涤），然后在37 ℃下用2XSSC、0.1%SDS洗涤两次1小时;放射自显影在-80 ℃下用筛网进行2-4天。参考文献：1. Jeffreys等人，1985，Nature 31A：67. 2. Jeffreys等人，1985年，《自然英国》：76。3. Burke等人，1987，Nature 27.：149. 4.达拉斯，1988，PNAS 8 j.：8014. 6. Feinberg，1984，Anal. 137：266. 7.约翰逊等人，1984年，吉恩·阿纳尔Tech.一比三。支持：NSF BSR-8700214（SHR）和BSR-8714397（BAS）; USDA PSW-84- 0021 CA（BAS）; NIH GM 31571（A.邓普顿）。图1：kb 9.4-
Some" DNA fingerprinting" probes are capable of revealing several moderately to hypervariable loci simultaneously when hybridized with blots of endonuclease-digested DNA from selected tetrapods (1, 2, 3). The 33.15 and 33.6 probes (1, 2) hybridize with sequences in rice (4); differences in hybridization patterns were found only among cultivars, but not among individuals within a cultivar. Here we show that probe 33.15 (supplied by A. Jeffreys; 1) reveals fragment differences between individuals from natural populations of the angiosperms Povulus deltoides (cottonwood; Fig. 1 lanes 1-6) and P. tremulodies (aspen; lanes 7, 8). In Fig. 1 unique numbers refer to different individuals. Lanes marked with a number and its prime represent two different branches fr'm the same individual, and similarity of such lanes implies that fragment lengths are somatically stable. While these results are preliminary, they demonstrate that probe 33.15 may hybridize with genomic sequences that are variable among individuals of dicot species, and with results from tetrapods (1, 2, 3) and monocots (4), suggest that particular minisatellite families (1, 2) may be widespread among the eukaryotes. Methods: 10 ug of DNA per individual extracted (5) from leaves was digested with HaeIII, electrophoresed in a 0.9% agofse gel, and blotted onto MSI Nylon. Probe 33.15 (1, 2) was hexamer-labelled (6) with P. Hybridization was in 1.4 XSSC, 0.5% nonfat dry milk solution (7) at 65 Cfor 12hr; filters were washed in IXDenhardts, 2XSSC, 0.2% SDS (two 15 min washes), then washed twice with 2XSSC, 0.1% SDS for I hr at 37 C; autoradiography was at-80 Cwith screens for 2-4 days. References: 1. Jeffreys et al., 1985, Nature 31A: 67. 2. Jeffreys et al., 1985, Nature Uk: 76. 3. Burke et al., 1987, Nature 27.: 149. 4. Dallas, 1988, PNAS 8j.: 8014. 6. Feinberg, 1984, Anal. Biochem. 137: 266. 7. Johnson et al., 1984, Gene Anal. Tech. 1: 3. Support: NSF BSR-8700214 (SHR) and BSR-8714397 (BAS); USDA PSW-84-0021CA (BAS); NIH GM31571 (A. Templeton). Fig. 1: kb 9.4-