US2005025787A1PendingUtilityA1

Transformed bacteria producing CS6 antigens as vaccines

Priority: May 13, 1994Filed: Jan 12, 2004Published: Feb 3, 2005
Est. expiryMay 13, 2014(expired)· nominal 20-yr term from priority
C07K 14/245A61P 37/04A61K 2039/523A61K 39/00
62
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Claims

Abstract

Disclosed herein are antigens that stimulate protective antibodies against enterotoxigenic Escherichia coli . Also disclosed herein are proteins encoded by cssA and cssB genes as well as constructs containing the genes and methods of using thereof.

Claims

exact text as granted — not AI-modified
1 . A method of inducing, in a susceptible host, the production of antibodies against CS6 protein comprising administration of a composition of matter comprising a bacteria transformed with a plasmid which contains genes  css A and  css B, all of the  css C and DNA sequence of  css D which encodes at least 802 amino acids (at least 2406 base pairs), an origin of replication, a Lac promotor, and a kanamycin resistance gene wherein said bacteria expresses both CS6A and CS6B proteins.  
     
     
         2 . A method of  claim 1  wherein the bacteria are in a pharmaceutically acceptable carrier.  
     
     
         3 . A method of  claim 2  wherein the carrier is a carbonated liquid.  
     
     
         4 . A protein containing the amino acid sequence  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                   RTEIATKNFPVSTTISKSFFA 
                     
                 
                     
                     
                 
                     
                   PEPRIQPSFGENVGKEGALLF 
                 
                     
                     
                 
                     
                   SVNLTVPENVSQVTVYPVYDE 
                 
                     
                     
                 
                     
                   DYGLGRLVNTADASQSIIYQI 
                 
                     
                     
                 
                     
                   VDEKGKKMLKDHGAEVTPNQQ 
                 
                     
                     
                 
                     
                   ITFKALNYTSGEKKISPGIYN 
                 
                     
                     
                 
                     
                   DQVMVGYYVN. 
                 
                     
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         5 . A composition of matter comprising a protein of  claim 4  in a pharmaceutically acceptable carrier.  
     
     
         6 . A bacteria transformed with a plasmid which contains genes  css A and  css B, all of the  css C and DNA sequence of  css D which encodes at least 802 amino acids (at least 2406 base pairs), an origin of replication, a Lac promotor, and a kanamycin resistance gene wherein said bacteria expresses both CS6A and CS6B proteins.  
     
     
         7 . A composition of matter comprising bacteria of  claim 6  in a pharmaceutically acceptable carrier.  
     
     
         8 . A composition of matter comprising the bacteria of  claim 6  wherein the carrier is a carbonated beverage.  
     
     
         9 . A bacteria of  claim 6  containing the sequence:  
       
         
           
                 
                 
                 
                 
               
                     
                 
                   1 
                   AAGCTTGTAA CCAGTTGATA AAAATATATC ACGCTGGGAA TGACGTGATG 
                   (Sequence  ID No  #1) 
                     
                 
                     
                 
                   51 
                   TATATACGGA GCAGCTATGT CGGAACAGAT ATTTTCCTAT CGGTATGCGT 
                 
                     
                 
                   101 
                   TGTGAGTAAG CGTAAAGCCA ATGCTGTCTG TAACTCCTGA TCCTTGCAGA 
                 
                     
                 
                   151 
                   CTAAATTAGA GCTCCTTCTA AATTAGACGG ATGGATAAAC CTACAGACTG 
                 
                     
                 
                   201 
                   GCGCTCTGGG TCTCGCCGGA TATTTTCTAA TGAATTTAAG CTTCATATGG 
                 
                     
                 
                   251 
                   TTGAACTGGC TTCGAAACCA AATGCCAATG TCGCACAACT GGCTCGGGAA 
                 
                     
                 
                   301 
                   CATGGCGTTG ATAACAACCT GATTTTTAAA TAGCTACGCC TCTGGCAAAG 
                 
                     
                 
                   351 
                   AGAAGGACGT ATTTCTCGTA GAATGCCTCC AACTATTGTA GGCCCTACAG 
                 
                     
                 
                   401 
                   TACCACTGAG GTAGCCTGAA TTTAAAGCCG AAGCGGTCAG AACTGTTCTT 
                 
                     
                 
                   451 
                   GGTGTGAACG TAGCACTCAC CAATAAAAGC ATCAATACGG TGCTCTGTTG 
                 
                     
                 
                   501 
                   ACACATTACG AATGTTATGT ATACAATAAA AATGATTATA GCAATATTAA 
                 
                     
                 
                   551 
                   TGGTGTTATA TGAAGAAAAC AATTGGTTTA ATTCTAATTC TTGCTTCATT 
                 
                     
                 
                   601 
                   CGGCAGCCAT GCCAGAACAG AAATAGCGAC TAAAAACTTC CCAGTATCAA 
                 
                     
                 
                   651 
                   CGACTATTTC AAAAAGTTTT TTTGCACCTG AACCACGAAT ACAGCCTTCT 
                 
                     
                 
                   701 
                   TTTGGTGAAA ATGTTGGAAA GGAAGGAGCT TTATTATTTA GTGTGAACTT 
                 
                     
                 
                   751 
                   AACTGTTCCT GAAAATGTAT CCCAGGTAAC GGTCTACCCT GTTTATGATG 
                 
                     
                 
                   801 
                   AAGATTATGG GTTAGGACGA CTAGTAAATA CCGCTGATGC TTCCCAATCA 
                 
                     
                 
                   851 
                   ATAATCTACC AGATTGTTGA TGAGAAAGGG AAAAAAATGT TAAAAGATCA 
                 
                     
                 
                   901 
                   TGGTGCAGAG GTTACACCTA ATCAACAAAT AACTTTTAAA GCGCTGAATT 
                 
                     
                 
                   951 
                   ATACTAGCGG GGAAAAAAAA ATATCTCCTG GAATATATAA CGATCAGGTT 
                 
                     
                 
                   1001 
                   ATGGTTGGTT ACTATGTAAA CTAAATACTG GAAGTATGAT TATGTTGAAA 
                 
                     
                 
                   1051 
                   AAAATTATTT CGGCTATTGC ATTAATTGCA GGAACTTCCG GAGTGGTAAA 
                 
                     
                 
                   1101 
                   TGCAGGAAAC TGGCAATATA AATCTCTGGA TGTAAATGTA AATATTGAGC 
                 
                     
                 
                   1151 
                   AAAATTTTAT TCCAGATATT GATTCCGCTG TTCGTATAAT ACCTGTTAAT 
                 
                     
                 
                   1201 
                   TACGATTCGG ACCCGAAACT GGATTCACAG TTATATACGG TTGAGATGAC 
                 
                     
                 
                   1251 
                   GATCCCTGCA GGTGTAAGCG CAGTTAAAAT CGCACCAACA GATAGTCTGA 
                 
                     
                 
                   1301 
                   CATCTTCTGG ACAGCAGATC GGAAAGCTGG TTAATGTAAA CAATCCAGAT 
                 
                     
                 
                   1351 
                   CAAAATATGA ATTATTATAT CAGAAAGGAT TCTGGCGCTG GTAACTTTAT 
                 
                     
                 
                   1401 
                   GGCAGGACAA AAAGGATCCT TTCCTGTCAA AGAGAATACG TCATACACAT 
                 
                     
                 
                   1451 
                   TCTCAGCAAT TTATACTGGT GGCGAATACC CTAATAGCGG ATATTCGTCT 
                 
                     
                 
                   1501 
                   GGTACTTATG CAGGAAATTT GACTGTATCA TTTTACAGCA ATTAAAAAAA 
                 
                     
                 
                   1551 
                   GGCCGCATTA TTGCGGCCAT TGACGATACT GCTAGGCAAA AATATGAAAT 
                 
                     
                 
                   1601 
                   CAAAGTTAAT TATACTATTG ACGTTAGTGC CATTTTCATC TTTTTCAACA 
                 
                     
                 
                   1651 
                   GGAAATAATT TTGAAATAAA TAAGACACGA GTAATTTACT CTGACAGCAC 
                 
                     
                 
                   1701 
                   ACCATCAGTT CAAATATCAA ATAATAAAGC ATATCCTTTA ATTATTCAAA 
                 
                     
                 
                   1751 
                   GCAATGTATG GGATGAAAGC AATAATAAAA ATCATGACTT TATAGCAACA 
                 
                     
                 
                   1801 
                   CCACCGATTT TTAAAATGGA AAGTGAAAGT CGGAATATAA TAAAAATAAT 
                 
                     
                 
                   1851 
                   TAAAACAACT ATTAATTTGC CGGACTCTCA GGAAAGTATG AGATGGTTAT 
                 
                     
                 
                   1901 
                   GTATTGAATC AATGCCACCA ATAGAAAAAA GTACTAAAAT AAACAGAAAA 
                 
                     
                 
                   1951 
                   GAAGGAAGGA CAGACAGTAT TAATATCAGC ATTCGGGGGT GCATTAAACT 
                 
                     
                 
                   2001 
                   GATATATCGA CCTGCCAGTG TTCCGTCTCC TGTTTTTAAT AATATAGTAG 
                 
                     
                 
                   2051 
                   AAAAATTAAA ATGGCATAAA AATGGAAAGT ATCTTGTATT AAAAAATAAT 
                 
                     
                 
                   2101 
                   ACACCCTATT ACATTAGCTT TTCTGAGGTT TTTTTTGATT CAGATAAAGT 
                 
                     
                 
                   2151 
                   AAACAATGCA AAAGATATTT TATATGTAAA ACCATACTCA GAGAAGAAAA 
                 
                     
                 
                   2201 
                   TAGATATCAG CAACAGAATA ATAAAAAAAA TCAAATGGGC TATGATTGAT 
                 
                     
                 
                   2251 
                   GATGCTGGCG CAAAAACAAA ACTTTATGAA TCAATTTTAT AAAAAATCTC 
                 
                     
                 
                   2301 
                   ATTACAGTAT ACAAAAACAT CAGATTACAG GCTTGCTTTT TTTGCTATTT 
                 
                     
                 
                   2351 
                   ATATATCCTT TCTCAACCTC ATATGGAAAT GAACAATTTA GTTTTGACTC 
                 
                     
                 
                   2401 
                   ACGATTCCTA CCATCAGGTT ATAATTACTC TTTAAATAGT AACTTACCTC 
                 
                     
                 
                   2451 
                   CTGAAGGTGA GTATCTGGTT GATATTTATA TTAACAAAAT AAAAAAGGAG 
                 
                     
                 
                   2501 
                   TCCGCGATTA TTCCTTTTTA TATAAAAGGA AATAAACTTG TACCATGTTT 
                 
                     
                 
                   2551 
                   ATCAAAAGAA AAAATTTCAT CTTTGGGTAT CAACATTAAT AATAACGACA 
                 
                     
                 
                   2601 
                   ACACAGAGTG TGTAGAAACA AGTAAGGCAG GTATTAGTAA TATCAGCTTT 
                 
                     
                 
                   2651 
                   GAGTTTAGCT CTCTTCGTTT GTTTATTGCT GTACCGAAAA ATCTTCTGTC 
                 
                     
                 
                   2701 
                   TGAGATTGAT AAAATATCAT CAAAGGATAT AGATAACGGG ATTCATGCTT 
                 
                     
                 
                   2751 
                   TATTTTTTAA TTATCAAGTA AATACAAGGC TAGCCAATAA TAAAAATCGT 
                 
                     
                 
                   2801 
                   TATGATTACA TTTCTGTTTC ACCAAATATA AATTATTTTT CATGGCGGTT 
                 
                     
                 
                   2851 
                   GCGTAATCTT TTTGAATTTA ACCAAAACAA CGATGAAAAA ACATGGGAAA 
                 
                     
                 
                   2901 
                   GAAACTACAC TTATCTAGAA AAAAGTTTTT ATGATAAAAA GCTAAACTTA 
                 
                     
                 
                   2951 
                   GTCGTTGGTG AAAGTTATAC GAATTCAAAT GTTTATAATA ACTACTCTTT 
                 
                     
                 
                   3001 
                   TACTGGTATT TCAGTTTCTA CAGATACAGA TATGTATACG CCAAGTGAAA 
                 
                     
                 
                   3051 
                   TCGATTATAC ACCAGAAATT CATGGAGTGG CTGATTCAGA CTCTCAGATT 
                 
                     
                 
                   3101 
                   ATTGTCAGGC AAGGCAACAC CATTATCATT AATGAAAGTG TTCCAGCCGG 
                 
                     
                 
                   3151 
                   ACCGTTCTCA TTTCCAATAA CCAATCTCAT GTATACTGGG GGGCAACTTA 
                 
                     
                 
                   3201 
                   ATGTGGAGAT AACAGATATT TATGGAAATA AAAAACAATA TACTGTCAAT 
                 
                     
                 
                   3251 
                   AATTCCTCTC TTCCTGTTAT GAGAAAAGCG GGACTAATGG TATATAATTT 
                 
                     
                 
                   3301 
                   TATATCTGGG AAATTAACAA AAAAAAATAG TGAGGATGGT GATTTTTTTA 
                 
                     
                 
                   3351 
                   CTCAAGGTGA TATTAACTAC GGTACTCACT ATAACAGCAC ACTATTCGGT 
                 
                     
                 
                   3401 
                   GGATATCAGT TTAGTAAAAA TTATTTTAAC TTATCTACTG GTATAGGCAC 
                 
                     
                 
                   3451 
                   TGATCTGGGA TTTTCTGGAG CATGGCTACT ACACGTTAGC AGAAGTAATT 
                 
                     
                 
                   3501 
                   TTAAGAATAA AAATGGATAT AATATTAATC TACAACAAAA CACTCAGTTA 
                 
                     
                 
                   3551 
                   AGACCATTCA ATGCCGGGGT TAATTTCGAT TACGCATACA GAAAAAAAAG 
                 
                     
                 
                   3601 
                   GTATGTGGAA CTTTCCGACA TTGGCTGGCA TGGTAATTTA TATAATCAAC 
                 
                     
                 
                   3651 
                   TTAAAAATAG TTTTTCTTTA TCCTTGTCAA AATCATTGAA TAAATACGGA 
                 
                     
                 
                   3701 
                   AATTTCTCAC TTGATTATAA CAAAATGAAA TACTGGGATA ATGCGTATGA 
                 
                     
                 
                   3751 
                   TAGTAACTCA ATGTCGATTC GTTATTTTTT TAAATTCATG CGAGCAATGA 
                 
                     
                 
                   3801 
                   TTACAACAAA TTGTTCTTTA AATAAATATC AATCTTATGA AAAAAAAGAT 
                 
                     
                 
                   3851 
                   AAAAGATTTA GTATTAATAT ATCATTGCCT TTAACCAAAG ATTACGGGCA 
                 
                     
                 
                   3901 
                   CATATCTTCA AACTATTCAT TTTCCAATGC AAATACACGA ACGGCAACCA 
                 
                     
                 
                   3951 
                   GTTCTGTAGG CTTAAACGGT AGTTTTTTTA ATGACGCAAG ATTAAACTGG 
                 
                     
                 
                   4001 
                   AACATTCAGC AGAACAGAAC GACCCGTAAC AATGGATATA CTGATAATAC 
                 
                     
                 
                   4051 
                   CAGTTACATA GCAACCAGCT ATGCCTCTCC CTATGGCGTT TTTACTGGTT 
                 
                     
                 
                   4101 
                   CATATTCAGG ATCGAACAAG TATTCAAGCC AGTTTTATTC TGCATCGGGA 
                 
                     
                 
                   4151 
                   GGTATTGTTT TGCATAGCGA TGGCGTAGCT TTTACTCAAA AAGCCGGAGA 
                 
                     
                 
                   4201 
                   TACCTCTGCT CTTGTCCGTA TTGATAATAT TTCTGATATA AAAATTGGTA 
                 
                     
                 
                   4251 
                   ACACTCCTGG TGTTTATACT GGGTATAATG GTTTTGCTTT AATTCCTCAT 
                 
                     
                 
                   4301 
                   CTTCAGCCGT TCAAAAAAAA CACCATTTTA ATTAATGATA AAGGAATTCC 
                 
                     
                 
                   4351 
                   AGACGGTATT ACTCTTGCTA ATATAAAAAA ACAAGTTATC CCATCACGAG 
                 
                     
                 
                   4401 
                   GAGCTATTGT TAAAGTAAAA TTTGATGCTA AAAAAGGCAA TGACATTTTG 
                 
                     
                 
                   4451 
                   TTTAAGCTTA CAACTAAAGA TGGAAAAACG CCCCCATTAG GAGCTATAGC 
                 
                     
                 
                   4501 
                   CCATGAAAAA AATGGAAAAC AGATTAATAC GGGTATCGTT GACGATGATG 
                 
                     
                 
                   4551 
                   GTATGCTTTA TATGTCTGGA TTATCAGGGA CAGGGATTAT TAATGTAACA 
                 
                     
                 
                   4601 
                   TGGAATGGAA AAGTCTGTTC ATTTCCTTTT TCAGAAAAAG ATATATCTAG 
                 
                     
                 
                   4651 
                   CAAACAATTA TCTGTTGTAA ATAAACAATG TTAGGTAGTG CATCCAATTA 
                 
                     
                 
                   4701 
                   GTAGAACATG TGTTTTTCGA TAAACGCTCC GATCTCTTTT TCGTGGATCT 
                 
                     
                 
                   4751 
                   CAACTGAGCG TGAGAAGCAG ATTGTTTTAC GAGCCAACCG CTTAATGCGG 
                 
                     
                 
                   4801 
                   GTGCGTAGCG TCAGATTATT ACGCTCAATG CGTTGGGTGA ATATTTTGCC 
                 
                     
                 
                   4851 
                   GGTCAGATGC TTATTCTTCG GTACC 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         10 . A protein expressed by the bacteria of  claim 6  containing the sequence:  
       
         
           
                 
                 
                 
                 
               
                     
                     
                 
                     
                   RTEIATKNFPVSTTISKSFFA 
                   (SEQ. ID No. 9) 
                     
                 
                     
                     
                 
                     
                   PEPRIQPSFGENVGKEGALLF 
                 
                     
                     
                 
                     
                   SVNLTVPENVSQVTVYPVYDE 
                 
                     
                     
                 
                     
                   DYGLGRLVNTADASQSIIYQI 
                 
                     
                     
                 
                     
                   VDEKGKKMLKDHGAEVTPNQQ 
                 
                     
                     
                 
                     
                   ITFKALNYTSGEKKISPGIYN 
                 
                     
                     
                 
                     
                   DQVMVGYYVN.

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