US2003166068A1PendingUtilityA1

Peptidoglycan recognition proteins and their production

Priority: Sep 17, 1996Filed: Apr 30, 2002Published: Sep 4, 2003
Est. expirySep 17, 2016(expired)· nominal 20-yr term from priority
Y10S530/855C07K 14/43586Y10S530/858
30
PatentIndex Score
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Claims

Abstract

A gene coding for a peptidoglycan recognition peptide (PGRP) is cloned, a recombinant vector into which said gene is introduced is obtained, and a transformant transformed with said recombinant vector is cultivated, thereby producing the PGRP in large amounts at high purity.

Claims

exact text as granted — not AI-modified
What is claimed is:  
     
         1 . A recombinant protein comprising the following amino acid sequence (SEQ ID NO: 1):  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   Asp Cys Asp Val Val Ser Lys Lys Gln Trp Asp Gly 
                     
                 
                     
                     
                 
                     
                   Leu Ile Pro Val His Val Ser Tyr Leu Ala Arg Pro 
                 
                     
                     
                 
                     
                   Val Ser Leu Val Ile Val Gln His Thr Val Thr Pro 
                 
                     
                     
                 
                     
                   Phe Cys Arg Thr Asp Ala Gly Cys Glu Glu Leu Val 
                 
                     
                     
                 
                     
                   Arg Asn Ile Gln Thr Asn His Met Glu Ala Leu Gln 
                 
                     
                     
                 
                     
                   Tyr Trp Asp Ile Gly Pro Ser Phe Leu Val Gly Gly 
                 
                     
                     
                 
                     
                   Asn Gly Lys Val Tyr Glu Gly Ser Gly Trp Leu His 
                 
                     
                     
                 
                     
                   Val Gly Ala His Thr Tyr Gly Tyr Asn Ser Arg Ser 
                 
                     
                     
                 
                     
                   Ile Gly Val Ala Phe Ile Gly Asn Phe Asn Thr Asp 
                 
                     
                     
                 
                     
                   Glu Pro Ser Gly Ala Met Leu Glu Ala Leu Arg Ser 
                 
                     
                     
                 
                     
                   Leu Leu Arg Cys Gly Val Glu Arg Gly His Leu Ala 
                 
                     
                     
                 
                     
                   Gly Asp Tyr Arg Val Val Ala His Arg Gln Leu Ile 
                 
                     
                     
                 
                     
                   Ala Ser Glu Ser Pro Gly Arg Lys Leu Tyr Asn Gln 
                 
                     
                     
                 
                     
                   Ile Arg Arg Trp Pro Glu Trp Leu Glu Asn Val Asp 
                 
                     
                     
                 
                     
                   Ser Ile Lys Asn Ala. 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         2 . The recombinant protein according to  claim 1 , wherein the recombinant protein has peptidoglycan recognition activity and has the following amino acid sequence (SEQ ID NO: 1) or one obtained by deletion, substitution or addition of one or more amino acids in the following amino acid sequence (SEQ ID NO: 1):  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   Asp Cys Asp Val Val Ser Lys Lys Gln Trp Asp Gly 
                     
                 
                     
                     
                 
                     
                   Leu Ile Pro Val His Val Ser Tyr Leu Ala Arg Pro 
                 
                     
                     
                 
                     
                   Val Ser Leu Val Ile Val Gln His Thr Val Thr Pro 
                 
                     
                     
                 
                     
                   Phe Cys Arg Thr Asp Ala Gly Cys Glu Giu Leu Val 
                 
                     
                     
                 
                     
                   Arg Asn Ile Gln Thr Asn His Met Glu Ala Leu Gln 
                 
                     
                     
                 
                     
                   Tyr Trp Asp Tle Gly Pro Ser Phe Leu Val Gly Giy 
                 
                     
                     
                 
                     
                   Asn Gly Lys Val Tyr Glu Gly Ser Gly Trp Leu His 
                 
                     
                     
                 
                     
                   Val Gly Ala His Thr Tyr Gly Tyr Asn Ser Arg Ser 
                 
                     
                     
                 
                     
                   Ile Gly Val Ala Phe Ile Gly Asn Phe Asn Thr Asp 
                 
                     
                     
                 
                     
                   Giu Pro Ser Gly Ala Met Leu Glu Ala Leu Arg Ser 
                 
                     
                     
                 
                     
                   Leu Leu Arg Cys Gly Val Glu Arg Gly His Leu Ala 
                 
                     
                     
                 
                     
                   Gly Asp Tyr Arg Val Val Ala His Arg Gln Leu Ile 
                 
                     
                     
                 
                     
                   Ala Ser Glu Ser Pro Gly Arg Lys Leu Tyr Asn Gin 
                 
                     
                     
                 
                     
                   Ile Arg Arg Trp Pro Glu Trp Leu Glu Asn Val Asp 
                 
                     
                     
                 
                     
                   Ser Ile Lys Asn Ala. 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         3 . A gene coding for a protein comprising the following amino acid sequence (SEQ ID NO: 1):  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   Asp Cys Asp Val Val Ser Lys Lys Gln Trp Asp Gly 
                     
                 
                     
                     
                 
                     
                   Leu Ile Pro Val His Val Ser Tyr Leu Ala Arg Pro 
                 
                     
                     
                 
                     
                   Val Ser Leu Val Ile Val Gln His Thr Val Thr Pro 
                 
                     
                     
                 
                     
                   Phe Cys Arg Thr Asp Ala Gly Cys Glu Glu Leu Val 
                 
                     
                     
                 
                     
                   Arg Asn Ile Gln Thr Asn His Met Glu Ala Leu Gln 
                 
                     
                     
                 
                     
                   Tyr Trp Asp Ile Gly Pro Ser Phe Leu Val Gly Gly 
                 
                     
                     
                 
                     
                   Asn Gly Lys Val Tyr Glu Gly Ser Gly Trp Leu His 
                 
                     
                     
                 
                     
                   Val Gly Ala His Thr Tyr Gly Tyr Asn Ser Arg Ser 
                 
                     
                     
                 
                     
                   Ile Gly Val Ala Phe Ile Gly Asn Phe Asn Thr Asp 
                 
                     
                     
                 
                     
                   Glu Pro Ser Gly Ala Met Leu Glu Ala Leu Arg Ser 
                 
                     
                     
                 
                     
                   Leu Leu Arg Cys Gly Val Glu Arg Gly His Leu Ala 
                 
                     
                     
                 
                     
                   Gly Asp Tyr Arg Val Val Ala His Arg Gln Leu Ile 
                 
                     
                     
                 
                     
                   Ala Ser Glu Ser Pro Gly Arg Lys Leu Tyr Asn Gln 
                 
                     
                     
                 
                     
                   Ile Arg Arg Trp Pro Glu Trp Leu Glu Asn Val Asp 
                 
                     
                     
                 
                     
                   Ser Ile Lys Asn Ala. 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         4 . The gene according to  claim 3 , wherein the protein has peptidoglycan recognition activity and has the following amino acid sequence (SEQ ID NO: 1) or one obtained by deletion, substitution or addition of one or more amino acids in the following amino acid sequence (SEQ ID NO: 1):  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   Asp Cys Asp Val Val Ser Lys Lys Gln Trp Asp Gly 
                     
                 
                     
                     
                 
                     
                   Leu Ile Pro Val His Val Ser Tyr Leu Ala Arg Pro 
                 
                     
                     
                 
                     
                   Val Ser Leu Val Ile Val Gln His Thr Val Thr Pro 
                 
                     
                     
                 
                     
                   Phe Cys Arg Thr Asp Ala Gly Cys Glu Glu Leu Val 
                 
                     
                     
                 
                     
                   Arg Asn Ile Gln Thr Asn His Net Glu Ala Leu Gln 
                 
                     
                     
                 
                     
                   Tyr Trp Asp Ile Gly Pro Ser Phe Leu Val Gly Gly 
                 
                     
                     
                 
                     
                   Asn Gly Lys Val Tyr Glu Gly Ser Gly Trp Leu His 
                 
                     
                     
                 
                     
                   Val Gly Ala His Thr Tyr Gly Tyr Asn Ser Arg Ser 
                 
                     
                     
                 
                     
                   Ile Gly Val Ala Phe Ile Gly Asn Phe Asn Thr Asp 
                 
                     
                     
                 
                     
                   Glu Pro Ser Gly Ala Met Leu Glu Ala Leu Arg Ser 
                 
                     
                     
                 
                     
                   Leu Leu Arg Cys Gly Val Glu Arg Gly His Leu Ala 
                 
                     
                     
                 
                     
                   Gly Asp Tyr Arg Val Val Ala His Arg Gln Leu Ile 
                 
                     
                     
                 
                     
                   Ala Ser Glu Ser Pro Gly Arg Lys Leu Tyr Asn Gln 
                 
                     
                     
                 
                     
                   Ile Arg Arg Trp Pro Glu Trp Leu Glu Asn Val Asp 
                 
                     
                     
                 
                     
                   Ser Ile Lys Asn Ala. 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         5 . A gene coding for a protein comprising the following amino acid sequence (SEQ ID NO: 2)  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   Met Ala Arg Leu His Ser Ala Val Val Leu Ala Leu 
                     
                 
                     
                     
                 
                     
                   Ala Leu Ser Ser Leu Leu Thr Glu Ile Ala Ala Asp 
                 
                     
                     
                 
                     
                   Cys Asp Val Val Ser Lys Lys Gln Trp Asp Gly Leu 
                 
                     
                     
                 
                     
                   Ile Pro Val His Val Ser Tyr Leu Ala Arg Pro Val 
                 
                     
                     
                 
                     
                   Ser Leu Val Ile Val Gln His Thr Val Thr Pro Phe 
                 
                     
                     
                 
                     
                   Cys Axg Thr Asp Ala Gly Cys Glu Glu Leu Val Arg 
                 
                     
                     
                 
                     
                   Asn Ile Gin Thr Asn His Met Glu Ala Leu Gln Tyr 
                 
                     
                     
                 
                     
                   Trp Asp Ile Gly Pro Ser Phe Leu Val Gly Gly Asn 
                 
                     
                     
                 
                     
                   Gly Lys Val Tyr Glu Gly Ser Gly Trp Leu His Val 
                 
                     
                     
                 
                     
                   Gly Ala His Thr Tyr Gly Tyr Asn Ser Arg Ser Ile 
                 
                     
                     
                 
                     
                   Gly Val Ala Phe Ile Gly Asn Phe Asn Thr Asp Glu 
                 
                     
                     
                 
                     
                   Pro Ser Gly Ala Met Leu Glu Ala Leu Arg Ser Leu 
                 
                     
                     
                 
                     
                   Leu Arg Cys Gly Val Glu Arg Gly His Leu Ala Gly 
                 
                     
                     
                 
                     
                   Asp Tyr Arg Val Val Ala His Arg Gln Leu Ile Ala 
                 
                     
                     
                 
                     
                   Ser Glu Ser Pro Gly Arg Lys Leu Tyr Asn Gin Ile 
                 
                     
                     
                 
                     
                   Arg Arg Trp Pro Glu Trp Leu Glu Asn Val Asp Ser 
                 
                     
                     
                 
                     
                   Ile Lys Asn Ala. 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         6 . The gene according to  claim 5 , wherein the protein has peptidoglycan recognition activity and has the following amino acid sequence (SEQ ID NO:2) or one obtained by deletion, substitution or addition of one or more amino acids in the following amino acid sequence (SEQ ID NO: 2):  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   Met Ala Arg Leu His Ser Ala Val Val Leu Ala Leu 
                     
                 
                     
                     
                 
                     
                   Ala Leu Ser Ser Leu Leu Thr Glu Ile Ala Ala Asp 
                 
                     
                     
                 
                     
                   Cys Asp Val Val Ser Lys Lys Gln Trp Asp Gly Leu 
                 
                     
                     
                 
                     
                   Ile Pro Val His Val Ser Tyr Leu Ala Arg Pro Val 
                 
                     
                     
                 
                     
                   Ser Leu Val Ile Val Gln His Thr Val Thr Pro Phe 
                 
                     
                     
                 
                     
                   Cys Arg Thr Asp Ala Gly Cys Glu Glu Leu Val Arg 
                 
                     
                     
                 
                     
                   Asn Ile Gln Thr Asn His Met Glu Ala Leu Gln Tyr 
                 
                     
                     
                 
                     
                   Trp Asp Ile Gly Pro Ser Phe Leu Val Gly Gly Asn 
                 
                     
                     
                 
                     
                   Gly Lys Val Tyr Glu Gly Ser Gly Trp Leu His val 
                 
                     
                     
                 
                     
                   Gly Ala His Thr Tyr Gly Tyr Asn Ser Arg Ser Ile 
                 
                     
                     
                 
                     
                   Gly Val Ala Phe Tle Gly Asn Phe Asn Thr Asp Glu 
                 
                     
                     
                 
                     
                   Pro Ser Gly Ala Met Leu Glu Ala Leu Arg Ser Leu 
                 
                     
                     
                 
                     
                   Leu Arg Cys Gly Val Glu Arg Gly His Leu Ala Gly 
                 
                     
                     
                 
                     
                   Asp Tyr Arg Val Val Ala His Arg Gln Leu Ile Ala 
                 
                     
                     
                 
                     
                   Ser Glu Ser Pro Gly Axg Lys Leu Tyr Asn Gln Tle 
                 
                     
                     
                 
                     
                   Arg Arg Trp Pro Glu Trp Leu Glu Asn Val Asp Ser 
                 
                     
                     
                 
                     
                   Ile Lys Asn Ala. 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         7 . A gene comprising a DNA having the following nucleotide sequence (SEQ ID NO: 3):  
       
         
           
                 
                 
               
                     
                 
                   GATTGCGACGTCGTCAGTAAAAAGCAATGGGACGGTTTGATCCCGGTGCA 
                     
                 
                     
                 
                   CGTGTCGTACCTGGCGCGGCCCGTGAGCCTCGTCATCGTCCAGCACACAG 
                 
                     
                 
                   TCACACCCTTCTGCAGGACGGACGCTGGCTGCGAGGAGCTCGTGCGTATC 
                 
                     
                 
                   CAGACCAACCACATGGAGGCCTTGCAATACTGGGACATCGGACCCTCGTT 
                 
                     
                 
                   CCTGGTGGGAGGTAACGGCAAGGTGTACGAGCGCTCCGGCTGCCTGCACG 
                 
                     
                 
                   TCGGCGCGCACACCTACGGGTACAACTCGACGTCCATCGGAGTCCATTCA 
                 
                     
                 
                   TCGGCAACTTCAACACGCACGACCCGAGCGGCGCGATGCTGGAGGCGCTG 
                 
                     
                 
                   CGGTCGCTGCTGCGCTGCGGCGTGGAGCGCGGCCACCTCGCGGGGGACTA 
                 
                     
                 
                   CCGCGTCGTGCCGCACCGACAGCTCATTGCCTCTGAGAGCCCCGGCCCGG 
                 
                     
                 
                   AACTCTACAACCAGATACGACGCTGGCCTGAGTGGCTGGAGAACGTGGAC 
                 
                     
                 
                   TCCATCAAGAACGCGTAA. 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         8 . A gene comprising a DNA including a nucleotide sequence which codes a protein having peptidoglycan recognition activity and is capable of hybridizing with the DNA having the following nucleotide sequence (SEQ ID NO: 3) under stringent conditions:  
       
         
           
                 
                 
               
                     
                 
                   GATTGCGACGTCGTCAGTAAAAAGCAATGGGACGGTTTGATCCCGGTGCA 
                     
                 
                     
                 
                   CGTGTCGTACCTGGCGCGGCCCGTGAGCCTCGTCATCGTCCAGCACACAG 
                 
                     
                 
                   TCACACCCTTCTGCAGGACGGACGCTGGCTGCGAGGAGCTCGTGCGGAAT 
                 
                     
                 
                   ATCCAGACCAACCACATGGAGGCCTTGCAATACTGGGACATCGGACCCTC 
                 
                     
                 
                   GTTCCTGGTGGGAGGTAACGGCAAGGTGTACGAGGGCTCCGGCTGGCTGC 
                 
                     
                 
                   ACGTCGGCGCGCACACCTACGGGTACAACTCGAGGTCCATCGGAGTCGCA 
                 
                     
                 
                   TTCATCGGCAACTTCAACACGGACGGCCGAGCGGCGCGATGCTGGAGGCG 
                 
                     
                 
                   CTGCGGTCGCTGCTGCGCTGCGGCGTGGAGCGCGGCCACCTCGCGGGGGA 
                 
                     
                 
                   CTACCGCGTCGTGGCGCACCGACAGCTCATTGCCTCTGAGAGCCCCGGCC 
                 
                     
                 
                   GGAAGCTCTACAAACCAGATACGACGCTGGCCTGAGTGGCTGGAGAACGT 
                 
                     
                 
                   GGACTCCATCAAGAACGCGTAA. 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         9 . A gene comprising a DNA having the following nucleotide sequence (SEQ ID NO: 5):  
       
         
           
                 
                 
               
                     
                 
                   ATGGCCCGCCTCCACTCGGCAGTTGTACTCGCGCTCGCTCTCAGCTCGCT 
                     
                 
                     
                 
                   TCTCACAGAAATAGCAGCCGATTGCGACGTCGTCAGTAAAAAGCAATGGG 
                 
                     
                 
                   ACGGTTTGATCCCGGTGCACGTGTCGTACCTGGCGCGGCCCGTGAGCCTC 
                 
                     
                 
                   GTCATCGTCCAGCACACAGTCACACCCTTCTGCAGGACGGACGCTGGCTG 
                 
                     
                 
                   CGAGCAGCTCGTGCGAATATCCAGACCAACCACATGGAGGCCTTGCAATA 
                 
                     
                 
                   CTGGGACATCGGACCCTCGTTCCTGGTGGGAGGTAACGGCAAGGTGTACG 
                 
                     
                 
                   AGGGCTCCGGCTGGCTGCACGTCGGCGCGCACACCTACGGGTACAACTCG 
                 
                     
                 
                   AGGTCCATCGGAGTCGCATTCATCGGCAACTTCAACACGGACGAGCCGAG 
                 
                     
                 
                   CGGCGCGATGCTGGAGGCGCTGCGGTCGCTGCTGCGCTGCGGCGTGGAGC 
                 
                     
                 
                   GCGGCCACCTCGCGGGGGACTACCGCGTCGTGGCGCACCGACAGCTCATT 
                 
                     
                 
                   GCCTCTGAGAGCCCCGGCCGGAACCTCTACAACCAGATACGACGCTGGCC 
                 
                     
                 
                   TGAGTGGCTGGAGAACGTGGACTCCATCAAGAACGCGTAA. 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         10 . A gene comprising a DNA including a nucleotide sequence which codes a protein having peptidoglycan recognition activity and is capable of hybridizing with the DNA having the following nucleotide sequence (SEQ ID NO: 5) under stringent conditions:  
       
         
           
                 
                 
               
                     
                 
                   ATGGCCCGCCTCCACTCGGCAGTTGTACTCGCGCTCGCTCTCAGCTCGCT 
                     
                 
                     
                 
                   TCTCACAGAAATAGCAGCCGATTGCGACGTCGTCAGTAAAAAGCAATGGG 
                 
                     
                 
                   ACGGTTTGATCCCGGTGCACGTGTCGTACCTGGCGCGGCCCGTGAGCCTC 
                 
                     
                 
                   GTCATCGTCCAGCACACAGTCACACCCTTCTGCAGGACGGACGCTGGCTG 
                 
                     
                 
                   CGAGAGCTCGTGCGATATCCAGACCAACCACATGGAGGCCTTGCAATACT 
                 
                     
                 
                   GGGACATCGGACCCTCGTTCCTGGTGGGAGGTAACGGCAAGGTGTACGAG 
                 
                     
                 
                   GGCTCCGGCTGGCTGCACGTCGGCGCGCACACCTACGGGTACAACTCGAC 
                 
                     
                 
                   GTCCATCGGAGTCGCATTCATCGGCAACTTCAACACGGACGAGCCGAGCG 
                 
                     
                 
                   GCGCGATGCTGGAGGCGCTGCGGTCGCTGCTGCGCTGCGGCGTGGAGCGC 
                 
                     
                 
                   GGCCACCTCGCGGGGGACTACCGCGTCGTGCGCACCGACAGCTCATTGCC 
                 
                     
                 
                   TCTGAGAGCCCCGGCCGGAAGCTCTACAACCAGATACGACGCTGGCCTGA 
                 
                     
                 
                   GTGGCTGGAGAACGTGGACTCCATCAAGAACGCGTAA. 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         11 . A recombinant vector comprising the gene of  claim 3 ,  5 ,  7  or  9 .  
     
     
         12 . A transformant transformed with the recombinant vector of  claim 11 .  
     
     
         13 . A method for producing a peptidoglycan recognition protein comprising cultivating the transformant of  claim 12  in a broth, and recovering the peptidoglycan recognition protein from the culture.

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