Some reports have indicated that the mutants of B. subtilis [36], Thermosynechococcus elongatus [37], and Borrelia burgdorferi [38] deficient in restriction endonuclease activity exhibited improved transformation efficiency.
H. salinarum strain Pho81Wr− (BR−HR−SRI−HtrI−SRII−HtrII−, carotenoid-deficient and restriction-deficient) (12) was used as the recipient in plasmid transformations.
On the other hand, LEPR allele Gln, which is deficient in MspI restriction site, was represented by a 212 bp fragment, whereas allele Arg, containing the restriction site, was represented by two bands of 151 and 61 bp in length.
P. putida KT2440 is a TOL plasmid cured, spontaneous restriction deficient derivative of P. putida mt-2 [ 3, 4].
In this work, we have developed and utilized a gene disruption plasmid (pSYCP) and intron enrichment strategy to generate a mutant strain of C. pasteurianum deficient in CpaAI restriction.
This includes RN6390, which is an 8325-4 strain in which three prophage were cured from NCTC 8325 [25], and SA113, which is a mutagenized, restriction-modification deficient derivative of 8325 [10], [26].
As the authors highlight in their article, hemodialysis may lead to a depletion of some biologically essential substances, possibly already deficient due to dietary restrictions.
These strains are derived from the restriction modification-deficient strain RM125 to possess a cI repressor gene cassette flanked by pBAC108-based BAC vector sequences [ 1, 10] and are identical with the exception that BEST6528 contains a 100 kb spacer sequence and an additional cI repressor gene cassette [ 16].
Mobilization by the conjugative plasmids was observed after short mating to other Neisseria species, to Haemophilus influenzae and to restriction-deficient Escherichia coli [27].
Briefly, J774 macrophages (about 4-5×104/well) seeded in 96-well culture plates were infected in triplicate with the transposon mutants, the non-polar deletion mutants, the restriction-deficient strain MFN245, or wild-type F. novicida U112 at the designated input MOI and washed at two hours post-infection (p.i).
