Jorgensen R A, Rothstein S J, Reznikoff W S
Mol Gen Genet. 1979;177(1):65-72. doi: 10.1007/BF00267254.
This paper reports a cleavage site map of Tn5 for restriction enzymes BamHI, Bg/I, Bg/II, Hind II, HindIII, HpaI, Sa/I, Aval, SmaI, XhoI, PstI, PvuII, HaeII and HaeIII that was determined by the analysis of restriction enzyme cleavage patterns of ColEl, two independent ColEl::Tn5 plasmids, and a ColEl::Tn5 deletion derivative. Ba/I, EcoRI, KpnI, and PvuI do not cleave Tn5. Construction and analysis of in vitro-generated deletions of a ColEl::Tn5 plasmid limit the sequences encoding neomycin resistance to a 1500-base-pair-long segment of Tn5. Insertion of DNA at a Bg/II site within this segment results in loss of the neomycin resistance phenotype. Since this Bg/II site lies in an inverted repeat region, sequences within this repeat seem to be involved in the expression of neomycin resistance.
本文报道了通过分析ColE1、两个独立的ColE1::Tn5质粒和一个ColE1::Tn5缺失衍生物的限制酶切割模式所确定的Tn5对于限制酶BamHI、Bg/I、Bg/II、Hind II、HindIII、HpaI、Sa/I、Aval、SmaI、XhoI、PstI、PvuII、HaeII和HaeIII的切割位点图谱。Ba/I、EcoRI、KpnI和PvuI不切割Tn5。对ColE1::Tn5质粒体外产生的缺失进行构建和分析，将编码新霉素抗性的序列限制在Tn5的一个1500碱基对长的片段内。在该片段内的Bg/II位点插入DNA会导致新霉素抗性表型的丧失。由于这个Bg/II位点位于一个反向重复区域内，该重复序列中的序列似乎参与了新霉素抗性的表达。
