ISSN:
1432-072X
Schlagwort(e):
Bacteriophage
;
Myxococcus
;
λ
;
Superooiled DNA
;
Cross-linking
;
Electron microscopy
Quelle:
Springer Online Journal Archives 1860-2000
Thema:
Biologie
Notizen:
Abstract DNA was partially released from the heads of myxococcus phages and also coliphage λ and examined by electron microscopy by a modification of the Kleinschmidt technique, in which water was used as hypophase. DNA emerged from the heads in patterns suggestive of newly relaxed supercoils. The unreleased DNA appeared to occupy discrete regions in the head. Some closed circles were released from λ heads. When NaCl solution was used as hypophase, the DNA was observed either released from the tail or from the head, in the latter case, supercoiled regions were observed. When NH4OAc solution was used as hypophase, tightly wound structures were released from λ heads; these fields also contained supercoiled circles. The presence of constrained supercoiled domains in newly released phage DNA was confirmed by observing the effects of ethidium bromide on its conformation. Treatment of phage with nitrogen mustard, a bifunctional alkylating agent, preserved supercoiled domains, even when the phage were lysed over water as hypophase. Further experiments suggested that phage inactivation by nitrogen mustard is largely due to restraint of the supercoiled, native, tertiary structure and that DNA-protein cross-linking may be involved in this reaction. The implications of these findings for the conformation of phage DNA in vivo are discussed and a new model for the winding of DNA in phage heads is proposed.
Materialart:
Digitale Medien
URL:
http://dx.doi.org/10.1007/BF00409932
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