Naslov
Transposon Tn5424 induces amplifications in Streptomyces rimosus

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

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

Izvornik
Universität Kaiserslautern VAAM Workshop "Biologie der Actinomyceten" / Cullum, J. - Kaiserslautern : Universität Kaiserslautern, 1998

Skup
VAAM Workshop "Biologie der Actinomyceten"

Mjesto i datum
Kaiserslautern, Njemačka, 04.10.1998. - 06.10.1998

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
Streptomyces; transposon; DNA amplification

Sažetak
Tn5424 is a transposon carrying the thiostrepton-resistance gene (tsr) based on the insertion sequence IS466 from Streptomyces coelicolor A3(2). It can be introduced into Streptomyces strains using derivatives of the temperature-sensitive replication plasmid pSG5. The original plasmid pUKG403 that transfers transposon Tn5424 was not convenient for use in Streptomyces rimosus because the plasmid-borne kanamycin resistance can not be selected in this species. In order to solve this problem an erythromycin resistance gene (mls) was introduced into the plasmid to give pPZG200. Selection at high temperature lead to loss of erythromycin resistance, but a certain proportion of the population retained the transposon thiostrepton-resistance marker indicating that transposition into the chromosome had occurred. When total DNA from these strains was examined by restriction analysis all turned out to carry high level DNA amplification of 1.5 kb and 9.0 kb in length. The amplified DNA hybridised to a tsr probe. The total DNA from all transposants hybridised to a cosmid clone pPZG25 (carrying the oxytetracycline cluster) showing the same pattern as in parental strain.

Izvorni jezik
Engleski

Znanstvena područja
Prehrambena tehnologija

POVEZANOST RADA