Rapid Characterization of S. mansoni Expression Library Clones of Potential Interest

Author:

KANAMURA Herminia Yohko1,HANCOCK Kathy2

Affiliation:

1. University of São Paulo

2. Centers for Disease Control and Prevention

Abstract

A S. mansoni adult worm cDNA expression library was screened with sera from baboons in a early phase after infection. The clones that were positive with the early infection sera were examined for reactivity with pre-infection sera and heterologous infection sera. In order to discriminate a positive antibody reaction from the reactivity due to residual anti-E. coli antibodies, an unrelated cDNA clone was plated with the positive clone. The unrelated clone provided the negative background and the contrast necessary to discern a positive antibody reaction. In this way, we were able to eliminate selected clones that were positive with the pre-infection sera or heterologous infection sera. This characterization of the expression library clones enabled us to quickly target only clones with the desired pattern of antibody reactivity for sequencing, subcloning, and expressing

Publisher

FapUNIFESP (SciELO)

Subject

Infectious Diseases,General Medicine

Reference4 articles.

1. Current protocols in molecular biology;AUSUBEL F. M.,1995

2. Development and optimization of the FAST-ELISA for detecting antibodies to Schistosoma mansoni;HANCOCK K.;J. immunol. Meth.,1986

3. Molecular cloning: a laboratory manual;SAMBROOK J.,1989

4. Optimization of the covalent conjugating procedure (NaIO4) of horseradish peroxidase to antibodies for use in enzyme-linked immunosorbent assay;TSANG V. C. W.;J. Immunoassay,1995

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