Avtor/Urednik		Gasparič, A; Nekrep, FV
Naslov		Ekspresijski sistemi za kloniranje genov anaerobnih vampnih bakterij
Prevedeni naslov		Expression systems for anaerobic rumen bacteria gene cloning
Tip		članek
Vir		Zb Biotehn Fak Univ Ljublj - Kmetijstvo
Vol. in št.		Letnik 62
Leto izdaje		1993
Obseg		str. 257-67
Jezik		slo
Abstrakt		By comparative experiments concerning the desired host-expression systems charactenstics, three of them: E. coli, B. subtilis and B. brevis 47-5 were tested. An expression system for cloning DNA fragments of anaerobic rumen bacteria coding for cellulases was selected. Chromosomal DNA sequence of Bacteroides succinogenes BL2 coding for endoglucanase was inserted in pNU200 plasmid in two orientations and used for B. subtilis MT119 transformation. Using different B. subtilis and E. coli strains we bypassed several problems connected with molecular genetics of anaerobic microorganisms. With recombinant vectors - cloning and expression plasmids pIL253 and pNU200 we tried to test a high secretory potenhal of B. brevis 47-5. Ruminococcus flavefaciens chromosomal DNA sequence CMC P 11 coding for endoglucanase was inserted in two orientations in plasmid pKK223-3. With this recombinant plasmid mixture the E. coli JM105 strain was transformed. The expression level was estimated in relation to insert orientation and different promoter usage.
Deskriptorji		ESCHERICHIA COLI BACILLUS SUBTILIS HYDROLYSIS GENE EXPRESSION REGULATION PLASMIDS CLONING, MOLECULAR