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...
Published in: | Revista do Instituto de Medicina Tropical de São Paulo |
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Universidade de São Paulo (USP)
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ftdoajarticles:oai:doaj.org/article:8b69482dac794999930d06b302ceda57 2024-09-09T19:27:14+00:00 Rapid Characterization of S. mansoni Expression Library Clones of Potential Interest Herminia Yohko KANAMURA Kathy HANCOCK 1997-11-01T00:00:00Z https://doi.org/10.1590/S0036-46651997000600009 https://doaj.org/article/8b69482dac794999930d06b302ceda57 EN eng Universidade de São Paulo (USP) http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46651997000600009 https://doaj.org/toc/0036-4665 https://doaj.org/toc/1678-9946 doi:10.1590/S0036-46651997000600009 0036-4665 1678-9946 https://doaj.org/article/8b69482dac794999930d06b302ceda57 Revista do Instituto de Medicina Tropical de São Paulo, Vol 39, Iss 6, p 355 (1997) cDNA Cloning Expression Lambda Schistosoma Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 article 1997 ftdoajarticles https://doi.org/10.1590/S0036-46651997000600009 2024-08-05T17:49:30Z 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 Uma biblioteca de expressão de cDNA de verme adulto de S. mansoni foi selecionada utilizando-se soros de macacos babuínos em fase inicial da infecção. Os clones que reagiram positivamente com os soros de infecção recente foram examinados quanto à reatividade contra soros normais e soros de infecção heteróloga. Com a finalidade de se conseguir melhor discriminação entre reatividade positiva com o anticorpo específico e aquela devido aos anticorpos anti-E. coli residuais, um clone de cDNA não relacionado ao S. mansoni foi plaqueado em mistura com o clone positivo. O plano de fundo negativo proporcionado pelo clone não relacionado forneceu o contraste necessário para discriminar a reação positiva específica. Esta metodologia de caracterização dos clones de expressão permite alcançar rapidamente os clones que apresentam o perfil de reatividade desejada, para posterior sequenciamento, subclonagem e expressão Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Lambda ENVELOPE(-62.983,-62.983,-64.300,-64.300) Plano ENVELOPE(-61.250,-61.250,-62.783,-62.783) Revista do Instituto de Medicina Tropical de São Paulo 39 6 355 359 |
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Open Polar |
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Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
cDNA Cloning Expression Lambda Schistosoma Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
spellingShingle |
cDNA Cloning Expression Lambda Schistosoma Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 Herminia Yohko KANAMURA Kathy HANCOCK Rapid Characterization of S. mansoni Expression Library Clones of Potential Interest |
topic_facet |
cDNA Cloning Expression Lambda Schistosoma Arctic medicine. Tropical medicine RC955-962 Infectious and parasitic diseases RC109-216 |
description |
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 Uma biblioteca de expressão de cDNA de verme adulto de S. mansoni foi selecionada utilizando-se soros de macacos babuínos em fase inicial da infecção. Os clones que reagiram positivamente com os soros de infecção recente foram examinados quanto à reatividade contra soros normais e soros de infecção heteróloga. Com a finalidade de se conseguir melhor discriminação entre reatividade positiva com o anticorpo específico e aquela devido aos anticorpos anti-E. coli residuais, um clone de cDNA não relacionado ao S. mansoni foi plaqueado em mistura com o clone positivo. O plano de fundo negativo proporcionado pelo clone não relacionado forneceu o contraste necessário para discriminar a reação positiva específica. Esta metodologia de caracterização dos clones de expressão permite alcançar rapidamente os clones que apresentam o perfil de reatividade desejada, para posterior sequenciamento, subclonagem e expressão |
format |
Article in Journal/Newspaper |
author |
Herminia Yohko KANAMURA Kathy HANCOCK |
author_facet |
Herminia Yohko KANAMURA Kathy HANCOCK |
author_sort |
Herminia Yohko KANAMURA |
title |
Rapid Characterization of S. mansoni Expression Library Clones of Potential Interest |
title_short |
Rapid Characterization of S. mansoni Expression Library Clones of Potential Interest |
title_full |
Rapid Characterization of S. mansoni Expression Library Clones of Potential Interest |
title_fullStr |
Rapid Characterization of S. mansoni Expression Library Clones of Potential Interest |
title_full_unstemmed |
Rapid Characterization of S. mansoni Expression Library Clones of Potential Interest |
title_sort |
rapid characterization of s. mansoni expression library clones of potential interest |
publisher |
Universidade de São Paulo (USP) |
publishDate |
1997 |
url |
https://doi.org/10.1590/S0036-46651997000600009 https://doaj.org/article/8b69482dac794999930d06b302ceda57 |
long_lat |
ENVELOPE(-62.983,-62.983,-64.300,-64.300) ENVELOPE(-61.250,-61.250,-62.783,-62.783) |
geographic |
Arctic Lambda Plano |
geographic_facet |
Arctic Lambda Plano |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
Revista do Instituto de Medicina Tropical de São Paulo, Vol 39, Iss 6, p 355 (1997) |
op_relation |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0036-46651997000600009 https://doaj.org/toc/0036-4665 https://doaj.org/toc/1678-9946 doi:10.1590/S0036-46651997000600009 0036-4665 1678-9946 https://doaj.org/article/8b69482dac794999930d06b302ceda57 |
op_doi |
https://doi.org/10.1590/S0036-46651997000600009 |
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Revista do Instituto de Medicina Tropical de São Paulo |
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39 |
container_issue |
6 |
container_start_page |
355 |
op_container_end_page |
359 |
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