Naslov
Characterisation of linear DNA ends in Streptomyces rimosus

Autori
Horvat, Lada Ivana ; Stoll, Alexander ; Hranueli, Daslav ; Cullum, John

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
Abstracts of the 99th General Meeting of the American Society for Microbiology / Maloney, P. - Chicago (IL) : American Society for Microbiology, 1999, 342 (H-66)

Skup
99th General Meeting of the American Society for Microbiology

Mjesto i datum
Chicago (IL), Sjedinjene Američke Države, 30.05.1999. - 03.06.1999

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Streptomyces rimosus; linear chromosome; linear plasmid; chromosomal and plasmid ends

Sažetak
S. rimosus has a linear chromosome of 8 Mb in size. In the "Zagreb strain" S. rimosus R6-65 there is a 320 kb linear plasmid pPZG102 which does not have any extensive terminal inverted repeats (Pandza et al., Mol. Microbiol., 28:1165, 1998). In a later derivative R6-500 in the strain development programme there is a 390 kb linear plasmid pPZG101 which has been derived from pPZG102 by the generation of long terminal repeats of about 180 kb and loss of about 110 kb of the pPZG102 sequences. Single cross overs between pPZG101 and the chromosome can generate chromosomes or plasmid primes carrying one chromosome end and one plasmid end. In order to study the structure of the plasmids in more detail it was necessary to characterise the ends. The ends of the linear chromosomes and linear plasmids in Streptomyces carry covalently bound proteins at the 5' ends. The ends can be isolated from DNA preparations using protein-binding materials such as glass milk (Huang et al., Mol Microbiol., 28:905, 1998). The isolation procedure was optimised to allow visualisation of terminal restriction fragments in ethidium-bromide stained agarose gels. In R6-65 three XhoI fragments of 4.7, 2.3 and 0.9 kb were observed. Southern hybridisation experiments identified the 4.7 kb fragment as the chromosome end. The other two fragments appear to correspond to the two ends of pPZG102. The lack of cross-hybridisation with the chromosome terminal fragment showed that despite the ability for stable replication of molecules carrying one chromosome and one plasmid end there is little conserved DNA homology between the ends.

Izvorni jezik
Engleski

Znanstvena područja
Prehrambena tehnologija

POVEZANOST RADA