Chemical linkage of the tail to the right-hand end of bacteriophage lambda DNA
Identifieur interne : 002012 ( Main/Exploration ); précédent : 002011; suivant : 002013Chemical linkage of the tail to the right-hand end of bacteriophage lambda DNA
Auteurs : John O. Thomas [États-Unis]Source :
- Journal of Molecular Biology [ 0022-2836 ] ; 1974.
Abstract
When solutions of bacteriophage λ are treated with a water-soluble carbodiimide (CMC), then lysed with formamide and observed with the electron microscope, over 90% of the DNA molecules are found to be attached to phage heads and tails. Sedimentation analysis shows that lysis is complete, and denaturation mapping shows that the right-hand end† † The right end of the DNA is defined as the (A + T)-rich end. of the DNA, and never the left-hand end, is attached to the phage tails. When the carbodiimide-treated phage are lysed and the DNA cleaved with RI endonuclease, the shortest fragment, which contains the right end of λ DNA, is as expected the only fragment found attached to the tails. Comparison of the length of the tail-linked fragment with the length of the free DNA fragment shows that the DNA does not extend to the end of the tail. It does appear to intrude a short distance (30% of the length of the tail, or 130 DNA base-pairs), but this distance is just at the limit of resolution. When phage are placed in 80% formamide and then immediately spread for electron microscopy, about 10% of the phage have partially ejected DNA. Denaturation mapping shows that it is the right end of the DNA which is ejected first.
Url:
DOI: 10.1016/0022-2836(74)90555-5
Affiliations:
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