Naslov
Homologous and heterologous expression of transfer RNA genes from Streptomyces rimosus

Autori
Durajlja, Sonja

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

Izvornik
Young Scientist's View of Molecular Biotechnology, Second International Meeting - FEBS Advanced Course, Strasbourg '93 / Joern, Putz ; Volker Doetsch - Strasbourg : Federation of European Biochemical Societies (FEBS), 1993, 43-43

Skup
Young Scientist's View of Molecular Biotechnology, Second International Meeting - FEBS Advanced Course, Strasbourg '93

Mjesto i datum
Strasbourg, Francuska, 28.06.1993

Vrsta sudjelovanja
Pozvano predavanje

Vrsta recenzije
Nije recenziran

Ključne riječi
Gene expression, promoters, transfer RNA genes, shuttle vectors, S. rimosus

Sažetak
Primary structure of seven tRNA genes from Streptomyces rimosus has been previously determined. Analysed genes include: two independent genes for tRNAfMet and the cluster containing two tRNAGln genes and three tRNAGlu genes. About twenty more tRNA genes from Streptomycin have also been analysed. Regions upstream from tRNA genes in Streptomyces show high degree of divergence and only few of sequenced genes contain promoter like elements. Sequences of regulatory elements responsible for the transcription of these genes are not known. In S. rimosus tRNAfMet1 gene and the cluster of Gln-Glu tRNA genes contain in the 5' flanking regions DNA sequences resembling to E.coli consensus promoter. The presumptive promoter of tRNAfMet2 gene show considerable sequence similarity in the -10 region to E.coli promoter, but there is no homology in -35 region. In this work, expression of S. rimosus tRNA genes has been studied in vivo in homologous system (S. rimosus) and in heterologous system (E.coli). All S. rimosus tRNA genes were (re)cloned in different combinations and orientations, with or without potential promoter regions, into stable bifunctional multi copy plasmid vectors pZG5 and pZG6, especially constructed for this purpose. Recombinant bifunctional plasmids with tRNA genes were introduced into both hosts. Total cellular tRNAs were isolated from S.rimosus and E.coli transform ants containing different recombinant plasmids and subjected to northern hybridisation analysis for the estimation of the relative amount of tRNAfMet, tRNAGln and tRNAGlu in different cell lines. Northern hybridisation analysis shows that all of analysed promoter are active in homologous system. From the deletion assay it can be concluded that all tRNA Gln and tRNAGlu genes are cotranscribed from the same promoter located 140-65 bp upstream from the first gene. Sequence of this promoter is TTGGAC-17-TAATGT. In the heterologous system (E.coli) only the cluster of Gln-Glu is properly expressed.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA