Restriction analysis of lambda EMBL3 background recombinants: occurrence of lambda phages carrying a head to tail oriented left arm DNA sequence |
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| Authors: | Helmut Tr?ster Wolfgang Kissel Michael F Trendelenburg and Ansgar Hofmann |
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| Institution: | (1) Institute of Experimental Pathology, German Cancer Research Center, Im Neuenheimer Feld 280, D-6900 Heidelberg, Federal Republic of Germany;(2) Institute of Toxicology, University of Mainz, Obere Zahlbacher Strasse 67, D-6500 Mainz, Federal Republic of Germany;(3) Present address: Boehringer GmbH, Postfach 310120, D-6800 Mannheim, Federal Republic of Germany |
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| Abstract: | Summary Eight representative recombinant background clones of λEMBL3 were analysed usingKpnI,BamHI,SalI,EcoRI andHindIII digestion. We found that λEMBL3 carries its own left arm in theBamHI cloning site. In this way, recombinant molecules were found to be generated which can grow onEscherichia coli strain NM539. In all cases analysed, the left arm DNA was inserted in a head to tail orientation. Seven clones carried a
restoredBamHI site at thecos site-BamHI site connection. In the region where the inserted left arm and the right arm were ligated,BamHI cloning produces a large palindromic sequence consisting of two polylinkers. ThisBamHI site was incompletely cleaved in all cases analysed. We assume that a part of the λ DNA molecule in this region shows a
cruciform structure prohibiting recognition or cleavage of this site by restriction endonucleaseBamHI. |
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| Keywords: | λ EMBL3 Recombinant background BamHI cloning Restriction endonuclease analysis |
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