title: Novel broad-host-range vehicles for cloning and shuffling of gene cassettes.
abstract: Novel vectors for cloning and shuffling of gene cassettes based on minireplicon of broad-host-range RA3 plasmid from IncU incompatibility group were constructed. A series of minireplicon variants were prepared with copy number ranging from low (1-2 copies per chromosome), medium (10-15 copies per chromosome) to high copy number (80-90 copies per chromosome). The new cloning vectors are relatively small in size (4.5-5.4kb) and carry various resistance determinants: kanamycin (Km(R)), tetracycline (Tc(R)) or chloramphenicol (Cm(R)). The vectors were engineered to facilitate cloning and shuffling of the functional modules with or without transcriptional terminators. Using the described strategy, a bank of functional modules, ready for exchange, has been initiated.
authors: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, 02-106 Warsaw, Pawinskiego 5A, Poland.
authors: Aneta A
authors: AA
authors: Bartosik
authors: Aleksandra
authors: A
authors: Markowska
authors: Jolanta
authors: J
authors: Szarlak
authors: Anna
authors: A
authors: Kulińska
authors: Grażyna
authors: G
authors: Jagura-Burdzy
mesh_terms: D001419:Bacteria / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D018628:Gene Dosage; D005822:Genetic Vectors / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D013045:Species Specificity