Pneumococcal surface protein C (PspC), epitopic regions and strain selection thereof, and uses therefor
a technology of surface protein and pneumococcal, applied in the field of pneumococcal surface protein c (pspc), can solve the problems of difficult approach, neonates and young children failing to make adequate immune responses to most, and achieving the effect of enhancing immunogenicity and enhancing immunogenicity
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[0098] Bacterial strains, plasmids, and recombinant DNA techniques
[0099] Chromosomal DNA from S. pneumoniae EF6796, a serotype 6A clinical isolate (Salser et al. 1993) and D39, a serotype 2 isolate, was isolated using a cesium chloride gradient procedure. The HindIII-EcoRI fragment of EF6796 and D39 was cloned in a modified pZero vector (Invitrogen, San Diego, Calif.) in which the Zeocin-resistance cassette was replaced by a kanamycin cassette, kindly provided by Randall Harris. Recombinant plasmids were electroporated into Escherichia coli TOP10F' cells [F'{lacI.sup.qTet.sup.R} mcrA_(mrr-hsdRMS-mcrBC) f80lacZ_M15_lacX74 deoR recA1 araD139_(ara-leu)7697 galU galK rpsL endA1 nupG] (Invitrogen). DNA was purified from agarose using Gene Clean (Bio101, Inc., Vista, Calif.).
[0100] Chromosomal DNA used for PCR was isolated using a chloroform-isoamyl alcohol procedure. Oligonucleotide primers, ABW13 (5' CGACGAATAGCTGAAGAGG 3') (SEQ ID NO: ) and SKH2 (5' CATA...
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