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[利用特异性核酸内切酶对噬菌体T5基因组的结构组织研究]

[Structural organization of the genome of bacteriophage T5 by specific endonucleases].

作者信息

Chernov A P, Kuz'min N P, Fodor I

出版信息

Mol Biol (Mosk). 1978 Nov-Dec;12(6):1329-40.

PMID:745606
Abstract

The DNA of Bacteriophage T5+ has been treated with restriction endonucleases EcoRI, HindIII, BamI, SmaI, PstI, SalI, KpnI and the electrophorectic 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 resolved from the electrophoretic pattern of double and partial digests, the digests of isolated restriction fragments and the digests of deletion mutant T5 st(O) DNA. Four BamI cleavage sites have been found and localized on the physical map of T5 DNA at 0.225; 0.68; 0.715 and 0.725 fractional lenght. Endonuclease PstI cuts T5 DNA at 11 sites: 0.095; 0.215; 0.320; 0.510; 0.635; 0.675; 0.710; 0.770; 0.810; 0.840; 0.875 fractional length. Six KpnI cleavage sites have been mapped at 0.140; 0.160; 0.530; 0.720; 0.760; 0.840 fractional length. 14 out of 17 HindIII cleavage sites have been localized at 0.10; 0.24; 0.255; 0.345; 0.40; 0.52; 0.54; 0.57; 0.70; 0.76; 0.80; 0.84; 0.87; 0.89, fractional length of T5 DNA. 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,以确定T5 DNA上的特异性切割位点。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T5 st(O) DNA的酶切图谱确定了切割位点的定位。已发现四个BamI切割位点,并将其定位在T5 DNA物理图谱上的0.225、0.68、0.715和0.725处(相对长度)。内切酶PstI在11个位点切割T5 DNA:0.095、0.215、0.320、0.510、0.635、0.675、0.710、0.770、0.810、0.840、0.875(相对长度)。六个KpnI切割位点已定位在0.140、0.160、0.530、0.720、0.760、0.840(相对长度)处。17个HindIII切割位点中的14个已定位在T5 DNA相对长度的0.10、0.24、0.255、0.345、0.40、0.52、0.54、0.57、0.70、0.76、0.80、0.84、0.87、0.89处。给出了七种限制性酶的噬菌体基因组完整切割图谱。

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