Method for sorting antifungal molecules acting on the glucanosyltransferase activity
AssigneeInstitut Pasteur
InventorsThierry Fontaine, Robbert Hartland, Isabelle Mouyna, Jean-Paul Latge
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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:
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 1-1913 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.