Restriction digests of the clone give the following sizes (kb): EcoRI--2.35, 2.35; HindIII--2.4, 2.3; SalI--4.7. 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 bacteria. pHP45:omega was cut with BssHII + SphI (removing the spcR/strR gene) and the ends were made blunt. This was ligated to a blunt-end 1.5 kb HindIII/SalI fragment of Tn5 containing the kanamycin resistance gene. |
