Restriction analysis of lambda EMBL3 background recombinants: occurrence of lambda phages carrying a head to tail oriented left arm DNA sequence.
Eight representative recombinant background clones of lambda EMBL3 were analysed using KpnI, BamHI, SalI, EcoRI and HindIII digestion. We found that lambda EMBL3 carries its own left arm in the BamHI cloning site. In the way, recombinant molecules were found to be generated which can grow on Escherichia coli strain NM539. In all cases analysed, the left arm DNA was inserted in a head to tail orientation. Seven clones carried a restored BamHI site at the cos 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. This BamHI site was incompletely cleaved in all cases analysed. We assume that a part of the lambda DNA molecule in this region shows a cruciform structure prohibiting recognition or cleavage of this site by restriction endonuclease BamHI.
Tröster
H
H
Institute of Experimental Pathology, German Cancer Research Center, Heidelberg, Federal Republic of Germany.
Kissel
W
W
Trendelenburg
M F
MF
Hofmann
A
A
D010582:Bacteriophage lambda / Q000235:genetics*; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D015245:Deoxyribonuclease BamHI / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D009690:Nucleic Acid Conformation
D004279:DNA, Viral; D015245:Deoxyribonuclease BamHI