Mobc (Mobilization protein C, mobc) (AP)

The mobilization region of the 7.4 kb beta-lactamase plasmid pSJ7.4 from Neisseria gonorrhoeae was characterized. The 3.2 kb HindIII-BamHI fragment of pSJ7.4 was mobilized between Escherichia coli strains by conjugative plasmid RK2. Selected restriction enzyme-generated deletions of this fragment were subcloned in pACYC177 to obtain constructs that were suitable for analysis of the mobilization region. Mating experiments showed that a highly conserved 1.9 kb DNA region within coordinates 4096-5997 is required for mobilization of pSJ7.4. This region contains two genes encoding the mobilization protein MobA, another protein, MobC, putatively involved in mobilization from some hosts, and an intergenic oriT. The 168 bp intergenic sequence also contains the promoters for mobA and mobC in an arrangement that suggests divergent transcription and autoregulation from oriT. The 56 kDa MobA was expressed in E. coli as a (6x)His-Tag fusion protein. Purified MobA specifically induced plasmid relaxation by nicking at the oriT. MobA is exceptional because the N-terminal region alone can mobilize pSJ7.4, albeit at a lower frequency than the full-length protein, even in the absence of MobC. The carboxyl terminal region of MobA did not share homology with other mobilization proteins, but may be involved in promoting efficient transfer of pSJ7.4.\n\nApplications:\nSuitable for use in Western Blot, ELISA.\n\nRecommended Dilution:\nELISA: 1:1000\nWestern Blot: 1:100-500\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C with the addition of glycerol. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. Do not freeze alkaline phosphatase conjugates which could result in a substantial loss of enzymatic activity. Light sensitive.

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SPECIFICATIONS

Catalog Number

031133-AP

Size

100ul

Applications

ELISA, WB

Hosts

Mouse

Form

Supplied as a liquid in PBS, pH 7.2. Contains ~0.09% sodium azide. Conjugated to Alkaline Phosphatase.

P Type

Mab

Purity

Purified by Protein G affinity chromatography.

Isotype

IgG2a

References

Rodriguez-Bonano,N.M.Plasmid 52 (2), 89-101 (2004) Rodriguez-Bonano,N.M.,Submitted (20-JAN-2003) Microbiology and Medical Zoology, Medical Sciences Campus, University of Puerto Rico, Puerto Rico Medical Center Area, Medical Sciences Main Building, Third Floor, Lab B-328, San Juan, PR 00936-5067, Puerto Rico

Additional Info

Neisseria gonorrhoeae

Alternative Names

Mobilization protein C, mobc