Size (kb): 4.4800000190734860
Vector: pHP45omega-Tc (plasmid)
Construction: pHP45:omega, pBR322 tetR
Construct size (kb): 4.480000019073486
Features: marker(s): ampR, tetR
terminator: T4D gene 32
vector permitting positive selection for integration
vector useful for interposon mutagenesis
Restriction digests of the clone give the following sizes (kb): EcoRI--2.5, 2.4; HindIII--2.45, 2.3; SalI--4.9.
Because resistance levels may vary, it is important to determine the proper antibiotic concentration for each strain or isolate.
One of a series (ATCC 37653
through 37657) of vectors for in vitro mutagenesis (via the omega interposon) of gram-negative genera.
pHP45:omega was cut with BssHII + SphI (removing the spcR/strR gene) and the ends were made blunt. This was ligated to a blunt-ended 1.4 kb HindIII/AvaI fragment of pBR322 containing the tetracycline resistance gene.
Fellay R, et al. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of Gram-negative bacteria. Gene 52: 147-154, 1987. PubMed: 3038679
Frey J, Krisch HM. Omega mutagenesis in Gram-negative bacteria: a selectable interposon which is strongly polar in a wide range of bacterial species. Gene 36: 143-150, 1985. PubMed: 2998930