Rapid characterization of S. mansoni expression library clones of potential interest.
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.
Kanamura
H Y
HY
School of Pharmaceutical Sciences, University of São Paulo, Brazil. kanamura@usp.br
Hancock
K
K
D000818:Animals; D002999:Clone Cells; D018509:DNA, Helminth / Q000235:genetics*; D015870:Gene Expression; D015723:Gene Library; D010215:Papio; D012550:Schistosoma mansoni / Q000235:genetics*
D018509:DNA, Helminth