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
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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-modifiedWhat 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.Join the waitlist — get patent alerts
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