OBTAINING AND STUDYING OF THE STREPTOMYCES GALILAEUS HKI022 EXCONJUGANTS
WHICH CARRY CONJUGATIVE PLASMIDS pWA1, pSET152 AND pWPK
A.Luzhetskyy, M.Fedoryshyn, V.Fedorenko
Ivan Franko National University of L'viv,
Hrushevskoho st.4, L'viv 79005, Ukraine,
e-mail: genetic@franko.lviv.ua
Plasmids pSET152 and pWA1 were transferred into strain Streptomyces galilaeus HKI022 (producer of aclacinomycins) with frequency 4,7 10-3 and 4,5 10-4 respectively. Eshcherichia coli DH1 (pUB307) and E.coli ET12567 (pUB307) were used as donors for mating. Integration of pSET152 into chromosome of S.galilaeus HKI022 has negative effects on antibiotic production. Hybridization analysis shows that two copies of pSET152 are integrated into chromosome of S.galilaeus HKI022. Plasmid pSET152 was inherited every time under non-selective conditions. S.galilaeus HKI022 (pWA1) exhibited a 100% stability of inheritance of the Thr marker after two passages, and 83% of clones inherited pWA1 after seven passages under non-selective conditions. Plasmid pWPK, which carries genes of cluster of landomycin E biosynthesis (lnd-cluster): lndA ( -ketoacylsynthase), lndB (chain lenght determinant), lndE (oxygenase) and lndD (ketoreductase), was constructed. Exconjugants S.galilaeus HKI022 (pWPK) were obtained with frequency 3,4 10-6. Spectrophotometric analysis of extracts from S.galilaeus HKI022 (pWPK) shows that this strain produces new compound with maximum of absorption 528 nm.
Key words: Streptomyces, conjugation, antitumor antibiotics.
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