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Libraries for two-hybrid screening of yeast and hyphal growth forms in Zymoseptoria tritici()
Pathogenic fungi are constantly emerging resistance to anti-fungal treatments. Therefore, identification of new fungicide targets is important. Good candidates are essential fungal proteins and their regulators. An efficient way to reveal the molecular environment of an essential protein is the sear...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Academic Press
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4502458/ https://www.ncbi.nlm.nih.gov/pubmed/26092795 http://dx.doi.org/10.1016/j.fgb.2015.03.023 |
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author | Ma, W. Kilaru, S. Collins, C. Courbot, M. Steinberg, G. |
author_facet | Ma, W. Kilaru, S. Collins, C. Courbot, M. Steinberg, G. |
author_sort | Ma, W. |
collection | PubMed |
description | Pathogenic fungi are constantly emerging resistance to anti-fungal treatments. Therefore, identification of new fungicide targets is important. Good candidates are essential fungal proteins and their regulators. An efficient way to reveal the molecular environment of an essential protein is the search for interacting factors. Here, we establish three yeast two-hybrid libraries, covering yeast and hyphal stages of the wheat pathogen Zymoseptoria tritici. No detectable genomic DNA was present in any of the 3 libraries. Random amplification revealed that the libraries include cDNA fragments of up to 2000 bp, suggesting that small-to-medium sized proteins are represented therein. Indeed, full-length cDNAs of five proteins were found in all libraries. The full-length cDNA of large chitin synthase gene mcs1 (5742 bp with introns; 5568 bp without introns) could not be amplified, but its 5′ and 3′ regions were represented, suggesting that even larger genes are covered in all libraries. Finally, we tested for the expected interaction of the autophagy proteins ZtAtg4 and ZtAtg8 in Z. tritici, and then used ZtAtg4 to screen one of the two-hybrid libraries. Indeed, we found ZtAtg8 as a positive interaction partner, confirming that interacting proteins can be identified. Thus, these molecular tools promise to be useful in identifying novel fungicide target proteins. |
format | Online Article Text |
id | pubmed-4502458 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Academic Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-45024582015-07-21 Libraries for two-hybrid screening of yeast and hyphal growth forms in Zymoseptoria tritici() Ma, W. Kilaru, S. Collins, C. Courbot, M. Steinberg, G. Fungal Genet Biol Article Pathogenic fungi are constantly emerging resistance to anti-fungal treatments. Therefore, identification of new fungicide targets is important. Good candidates are essential fungal proteins and their regulators. An efficient way to reveal the molecular environment of an essential protein is the search for interacting factors. Here, we establish three yeast two-hybrid libraries, covering yeast and hyphal stages of the wheat pathogen Zymoseptoria tritici. No detectable genomic DNA was present in any of the 3 libraries. Random amplification revealed that the libraries include cDNA fragments of up to 2000 bp, suggesting that small-to-medium sized proteins are represented therein. Indeed, full-length cDNAs of five proteins were found in all libraries. The full-length cDNA of large chitin synthase gene mcs1 (5742 bp with introns; 5568 bp without introns) could not be amplified, but its 5′ and 3′ regions were represented, suggesting that even larger genes are covered in all libraries. Finally, we tested for the expected interaction of the autophagy proteins ZtAtg4 and ZtAtg8 in Z. tritici, and then used ZtAtg4 to screen one of the two-hybrid libraries. Indeed, we found ZtAtg8 as a positive interaction partner, confirming that interacting proteins can be identified. Thus, these molecular tools promise to be useful in identifying novel fungicide target proteins. Academic Press 2015-06 /pmc/articles/PMC4502458/ /pubmed/26092795 http://dx.doi.org/10.1016/j.fgb.2015.03.023 Text en © 2015 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Ma, W. Kilaru, S. Collins, C. Courbot, M. Steinberg, G. Libraries for two-hybrid screening of yeast and hyphal growth forms in Zymoseptoria tritici() |
title | Libraries for two-hybrid screening of yeast and hyphal growth forms in Zymoseptoria tritici() |
title_full | Libraries for two-hybrid screening of yeast and hyphal growth forms in Zymoseptoria tritici() |
title_fullStr | Libraries for two-hybrid screening of yeast and hyphal growth forms in Zymoseptoria tritici() |
title_full_unstemmed | Libraries for two-hybrid screening of yeast and hyphal growth forms in Zymoseptoria tritici() |
title_short | Libraries for two-hybrid screening of yeast and hyphal growth forms in Zymoseptoria tritici() |
title_sort | libraries for two-hybrid screening of yeast and hyphal growth forms in zymoseptoria tritici() |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4502458/ https://www.ncbi.nlm.nih.gov/pubmed/26092795 http://dx.doi.org/10.1016/j.fgb.2015.03.023 |
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