Use of rpsL as a Counterselectable Marker in Borrelia burgdorferi.
نویسندگان
چکیده
We have demonstrated that rpsL, encoding the S12 protein of the small ribosomal subunit, can be used as a counterselectable marker in Borrelia burgdorferi, the causative agent of Lyme disease. Mutations in rpsL confer streptomycin resistance. Streptomycin susceptibility is dominant in an rpsL merodiploid, and streptomycin selects for the loss of wild-type rpsL carried in trans. This is the first description of a counterselectable marker in B. burgdorferi.
منابع مشابه
Modulation of Borrelia burgdorferi stringent response and gene expression during extracellular growth with tick cells.
Borrelia burgdorferi N40 multiplied extracellularly when it was cocultured with tick cells in L15BS medium, a medium which by itself did not support B. burgdorferi N40 growth. Growth of B. burgdorferi N40 in the presence of tick cells was associated with decreased production of (p)ppGpp, the stringent response global regulator, a fourfold decrease in relA/spoT mRNA, an eightfold net decrease in...
متن کاملIdentification of Two Epitopes on the Outer Surface Protein A of the Lyme Disease Spirochete Borrelia burgdorferi
A murine IgM monoclonal antibody (MA-2C6) with κ-light chains directed against an antigenic determinant of outer surface protein A (OspA) of the Lyme disease spirochete, Borreliaburgdorferi, is produced. This antibody could bind specifically to OspA antigen of several isolates of B. burgdorferi, but not to the non-Lyme disease bacteria such as T. pallidum and B. hermsii. Antibody MA-2C6 was pur...
متن کاملExpression and Purification of Recombinant Outer Surface Protein D of Borrelia burgdorferi
To carry out the immunological experiments on the serum of Multiple Sclerosis (MS) patients, based on a correlation between Borrelia burgdorferi infection and contracting MS autoimmune disease the outer surface protein D (OspD) of the bacterium was expressed and purified. A clone containing the OspD gene in pET11a expression vector under the control of T7 promoter was transformed to the bacteri...
متن کاملEfficient targeted mutagenesis in Borrelia burgdorferi.
Genetic studies in Borrelia burgdorferi have been hindered by the lack of a nonborrelial selectable marker. Currently, the only selectable marker is gyrB(r), a mutated form of the chromosomal gyrB gene that encodes the B subunit of DNA gyrase and confers resistance to the antibiotic coumermycin A(1). The utility of the coumermycin-resistant gyrB(r) gene for targeted gene disruption is limited b...
متن کاملAn rpsL cassette, janus, for gene replacement through negative selection in Streptococcus pneumoniae.
Natural genetic transformation offers a direct route by which synthetic gene constructs can be placed into the single circular chromosome of Streptococcus pneumoniae. However, the lack of a general negative-selection marker has hampered the introduction of constructs that do not confer a selectable phenotype. A 1.3-kb cassette was constructed comprising a kanamycin (Kn) resistance marker (kan) ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Applied and environmental microbiology
دوره 76 3 شماره
صفحات -
تاریخ انتشار 2010