Method for sorting antifungal molecules acting on the glucanosyltransferase activity
AssigneeInstitut Pasteur
InventorsThierry Fontaine, Robbert Hartland, Isabelle Mouyna, Jean-Paul Latge
Life of this patent
Estimating the term…
Abstract
The invention discloses proteins having β(1-3)glucanosyltransferase type activities. These proteins can be used for detecting the antifungal activity of molecules.
Claims23
Protein having an activity of the β-(1-3)-glycanosyltransferase type characterized in that it has at least 50%, preferably 60%, and even more preferably 85% homology with the protein having one of the sequences, or at least a part of the sequences SEQ ID No 2 or SEQ ID No 3 as follows:
- 8 Met Lys Ala Ser Ala Val Thr Ala Ala Leu Ala Val Gly Ala Ser Thr SEQ ID NO:2 Val Leu Ala Ala Pro Ser Ile Lys Ala Arg Asp Asp Val Thr Pro Ile Thr Val Lys Gly Asn Ala Phe Phe Lys Gly Asp Glu Arg Phe Tyr Ile Arg Gly Val Asp Tyr Gln Pro Gly Gly Ser Ser Asp Leu Ala Asp Pro Ile Ala Asp Ala Asp Gly Cys Lys Arg Asp Ile Ala Lys Phe Lys Glu Leu Gly Leu Asn Thr Ile Arg Val Tyr Ser Val Asp Asn Ser Lys Asn His Asp Glu Cys Met Asn Thr Leu Ala Asp Ala Gly Ile Tyr Leu Val Leu Asp Val Asn Thr Pro Lys Tyr Ser Ile Asn Arg Ala Lys Pro Lys Glu Ser Tyr Asn Asp Val Tyr Leu Gln Tyr Ile Phe Ala Thr Val Asp Ala Phe Ala Gly Tyr Lys Asn Thr Leu Ala Phe Phe Ser Gly Asn Glu Val Ile Asn Asp Gly Pro Ser Ser Ser Ala Ala Pro Tyr Val Lys Ala Val Thr Arg Asp Leu Arg Gln Tyr Ile Arg Ser Arg Lys Tyr Arg Glu Ile Pro Val Gly Tyr Ser Ala Ala Asp Ile Asp Thr Asn Arg Leu Gln Met Ala Gln Tyr Met Asn Cys Gly Ser Asp Asp Glu Arg Ser Asp Phe Phe Ala Phe Asn Asp Tyr Ser Thr Cys Asp Pro Ser Ser Phe Lys Thr Ser Gly Trp Asp Gln Lys Val Lys Asn Phe Thr Gly Tyr Gly Leu Pro Leu Phe Leu Ser Glu Tyr Gly Cys Asn Thr Asn Lys Arg Gln Phe Gln Glu Val Ser Ser Leu Tyr Ser Thr Asp Met Thr Gly Val Tyr Ser Gly Gly Leu Val Tyr Glu Tyr Ser Gln Glu Ala Ser Asn Tyr Gly Leu Val Glu Ile Ser Gly Asn Asn Val Lys Glu Leu Pro Asp Phe Asp Ala Leu Lys Thr Ala Phe Glu Lys Thr Ser Asn Pro Ser Gly Asp Gly Asn Tyr Asn Lys Thr Gly Gly Ala Asn Pro Cys Pro Ala Lys Asp Ala Pro Asn Trp Asp Val Asp Asn Asp Ala Leu Pro Ala Ile Pro Glu Pro Ala Lys Lys Tyr Met Thr Glu Gly Ala Gly Lys Gly Pro Gly Phe Ala Gly Pro Gly Ser Gln Asp Arg Gly Thr Gln Ser Thr Ala Thr Ala Glu Pro Gly Ser Gly Ser Ala Thr Gly Ser Ser Ser Ser Gly Thr Ser Thr Ser Ser Lys Gly Ala Ala Ala Gly Leu Thr Val Pro Ser Leu Thr Met Ala Pro Val Val Val Gly Ala Val Thr Leu Leu Ser Thr Val Phe Gly Ala Gly Leu Val Leu Leu:
- Asp Asp Val Thr Pro Ile SEQ ID NO:3 Thr Val Lys Gly Asn Ala Phe Phe Lys Gly Asp Glu Arg Phe Tyr Ile Arg Gly Val Asp Tyr Gln Pro Gly Gly Ser Ser Asp Leu Ala Asp Pro Ile Ala Asp Ala Asp Gly Cys Lys Arg Asp Ile Ala Lys The Lys Glu Leu Gly Leu Asn Thr Ile Arg Val Tyr Ser Val Asp Asn Ser Lys Asn His Asp Glu Cys Met Asn Thr Leu Ala Asp Ala Gly Ile Tyr Leu Val Leu Asp Val Asn Thr Pro Lys Tyr Ser Ile Asn Arg Ala Lys Pro Lys Glu Ser Tyr Asn Asp Val Tyr Leu Gln Tyr Ile Phe Ala Thr Val Asp Ala Phe Ala Gly Tyr Lys Asn Thr Leu Ala Phe Phe Ser Gly Asn Gln Val Ile Asn Asp Gly Pro Ser Ser Ser Ala Ala Pro Tyr Val Lys Ala Val Thr Arg Asp Leu Arg Gln Tyr Ile Arg Ser Arg Lys Tyr Arg Gln Ile Pro Val Gly Tyr Ser Ala Ala Asp Ile Asp Thr Asn Arg Leu Gln Met Ala Gln Tyr Met Asn Cys Gly Ser Asp Asp Gln Arg Ser Asp Phe Phe Ala Phe Asn Asp Tyr Ser Trp Cys Asp Pro Ser Ser Phe Lys Thr Ser Gly Trp Asp Gln Lys Val Lys Asn Phe Thr Gly Tyr Gly Leu Pro Leu Phe Leu Ser Gln Tyr Gly Cys Asn Thr Asn Lys Arg Gln Phe Gln Glu VaT Ser Ser Leu Tyr Ser Thr Asp Met Thr Gly Val Tyr Ser Gly Gly Leu Val Tyr Glu Tyr Ser Gln Gln Ala Ser Asn Tyr Gly Leu Val Gln Ile Ser Gly Asn Asn Val Lys Glu Leu Pro Asp Phe Asp Ala Leu Lys Thr Ala Phe Glu Lys Thr Ser Asn Pro Ser Gly Asp Gly Asn Tyr Asn Lys Thr Gly Gly Ala Asn Pro Cys Pro Ala Lys Asp Ala Pro Asn Thr Asp Val Asp Asn Asp Ala Leu Pro Ala Ile Pro Glu Pro Ala Lys Lys Tyr Met Thr Gln Gly Ala Gly Lys Gly Pro Gly Phe Ala Gly Pro Gly Ser Gln Asp Arg Gly Thr Gln Ser Thr Ala Thr Ala Glu Pro Gly Ser Gly Ser Ala Thr Gly Ser Ser Ser Ser Gly Thr Ser Thr Ser Ser Lys Gly Ala Ala Ala Gly Leu Thr Val Pro Ser Leu Thr Met Ala Pro Val Val Val Gly Ala Val Thr Leu Leu Ser Thr Val Phe Gly Ala Gly Leu Val Leu Leu:
Protein having an activity of the β-(1-3)-glycanosyltransferase type characterized in that it has at least 50%, preferably 60% and even more preferably 85% homology with the protein having the sequence SEQ ID No 10, or at least a part of the sequence SEQ ID No 10 as follows:
- 9 Gly Phe Phe Ala Gly Asn Glu Val Ile Asn Glu Gln Ser Val Lys Asn SEQ ID NO 10 Val Pro Thr Tyr Val Arg Val Cys His Pro Ser Pro Gln Leu Thr Ile Ala Cys Pro Leu:
Protein having an activity of the β-(1-3)-glycanosyltransferase type characterized in that it has at least 50%, preferably 60% and even more preferably 85% homology with the protein having the sequence SEQ ID No 12, or at least a part of the sequence SEQ ID No 12 as follows:
- 10 (BGT4) Gly Phe Phe Ala Gly Asn Glu Val Val Asn Gln Ala Asn Gln Ser Ala SEQ ID NO 12 Gly Ala Ala Phe Val Lys Ala Ala Ala Arg Asp Met Lys Ala Tyr Ile Lys Thr Lys Gly Tyr Arg Gln Ser Leu Ala Ile Gly Tyr Ala Thr Thr Asp Asn Pro Glu Ile Arg Leu Pro Leu Ser Asp Tyr Leu Asn Cys Gly Asp Gln Ala Asp Ala Val Asp Phe Phe Gly Tyr Asn Ile Tyr Glu Trp Cys Gly Asp Gln Thr Phe Gln Thr Ser Gly Tyr Gln Asn Arg Thr Glu Glu Tyr Lys Asp Tyr Ser Ile Pro Ile Phe Ile Ser Glu Tyr Gly Cys Asn:
E. coli strain deposited at the CNCM under the deposit number I-1762 carrying the sequence SEQ ID No 1.
E coli strain deposited at the CNCM under the deposit number I-1914 carrying the sequence SEQ ID No 9.
E. coli strain deposited at the CNCM under the deposit number I-1913 carrying the sequence SEQ ID No 11.
Method for detecting a nucleotide sequence having at least 60% identity with the sequence SEQ ID No 1, in a biological sample containing nucleotide sequences, comprising the following steps:
- a) placing the biological sample in contact with the nucleotide primers P3 and P4 having the sequences SEQ ID No 7 and SEQ ID No 8, respectively, as follows:
- 14 GSYTTCTTCK CYGGCAACGA GGTT:
- SEQ ID No7 GTTGCAGCCG WATTCGGASA YGAA:
- SEQ ID No8
- the nucleotide sequences contained in the sample having been if necessary put into a form enabling their hybridization under conditions enabling the hybridization of the primers with the nucleotide sequences.
- b) amplification of the nucleotide sequences
- c) revealing the amplification products, and
- d) detection of the mutations by appropriate methods.
Dependencies are inferred from the claim text.