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噬菌体T5基因组上限制性内切核酸酶识别的切割位点的物理图谱。

Physical mapping of cleavage sites recognized by restriction endonucleases on the genome of bacteriophage T5.

作者信息

Tchernov A P, Kouzmin N P, Fodor I

出版信息

Gene. 1978 Jul;3(4):293-302. doi: 10.1016/0378-1119(78)90039-2.

Abstract

The DNA of bacteriophage T5 has been treated with restriction endonucleases EcoRI, HindIII, BamI, SmaI, PstI, SalI, KpnI and the electrophoretic pattern obtained in agarose gel has been analyzed in order to localize the specific cleavage sites on the T5 DNA. The localization of cleavage sites has been reduced from the electrophoretic pattern of double and partial digests, the digests of isolated restriction fragments and the digests of deletion mutant T5st(o) DNA. Four BamI cleavage sites have been found and localized on the physical map of T5 DNA at 0.21, 0.225, 0.685 and 0.725 fractional length. Endonuclease SmaI cleaves at 0.39, 0.59 and 0.69 fractional length. Endonuclease PstI cuts T5 DNA at 11 sites: 0.090, 0.210, 0.320, 0.510, 0.635, 0.670, 0.705, 0.770, 0.815, 0.840, 0.875 fractional length. Six KpnI cleavage sites have been mapped at 0.170, 0.215, 0.525, 0.755, 0.830, 0.850 fractional length. A complete cleavage map of the phage genome is presented for seven restriction enzymes.

摘要

用限制性内切酶EcoRI、HindIII、BamI、SmaI、PstI、SalI、KpnI处理噬菌体T5的DNA,并分析在琼脂糖凝胶中获得的电泳图谱,以便确定T5 DNA上的特定切割位点。通过双酶切和部分酶切的电泳图谱、分离的限制性片段的酶切图谱以及缺失突变体T5st(o) DNA的酶切图谱,确定了切割位点的位置。已发现四个BamI切割位点,并将其定位在T5 DNA物理图谱上,分别位于0.21、0.225、0.685和0.725的分数长度处。内切酶SmaI在0.39、0.59和0.69的分数长度处切割。内切酶PstI在11个位点切割T5 DNA:0.090、0.210、0.320、0.510、0.635、0.670、0.7

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