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利用限制性核酸酶和分子克隆技术对非洲绿猴细胞α-卫星DNA的组织进行的研究。

Studies on the organization of the alpha-satellite DNA from African green monkey cells using restriction nucleases and molecular cloning.

作者信息

Graf H, Fittler F, Zachau H G

出版信息

Gene. 1979 Feb;5(2):93-110. doi: 10.1016/0378-1119(79)90096-9.

Abstract

alpha-Satellite DNA from African green monkey cells was analysed with restriction nucleases in some detail confirming and complementing our earlier results. With EcoRI and HaeIII (or BsuRI isoschizomer), about 25 and 10%, respectively, of the satellite DNA were cleaved into a series of fragments of the 172 bp repeat length and multiples thereof. To allow studies with fragments of homogeneous sequence unit length, HindIII fragments were covalently joined with the plasmid pBR 313. After transformation 19 clones were obtained, containing up to three monomer fragments. Nine of the clones were characterized by digestion with EcoRI. Three of these had cleavage sites for this nuclease in the satellite DNA portion. In the six clones tested with HaeIII no cleavage site was detected in the cloned DNA. The results are discussed in relation to the nucleotide sequence data recently published by Rosenberg et al. (1978) and in the context of random and nonrandom processes in satellite DNA evolution

摘要

我们对非洲绿猴细胞中的α卫星DNA进行了较为详细的限制性核酸酶分析,证实并补充了我们之前的研究结果。使用EcoRI和HaeIII(或BsuRI同裂酶)时,分别约有25%和10%的卫星DNA被切割成一系列长度为172 bp的重复片段及其倍数片段。为了能够对具有均匀序列单元长度的片段进行研究,将HindIII片段与质粒pBR 313共价连接。转化后获得了19个克隆,其中包含多达三个单体片段。通过用EcoRI消化对9个克隆进行了表征。其中三个在卫星DNA部分具有该核酸酶的切割位点。在用HaeIII测试的六个克隆中,未在克隆DNA中检测到切割位点。结合Rosenberg等人(1978年)最近发表的核苷酸序列数据以及卫星DNA进化中的随机和非随机过程对结果进行了讨论。

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