Nucleic acid sequences encoding two-component sensing and regulatory proteins, antimicrobial proteins and uses therefor
AssigneeNorth Carolina State University
InventorsTodd Klaenhammer, William Russell, Eric Altermann
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Abstract
Stress-related nucleic acid molecules and polypeptides and fragments and variants thereof are disclosed in the current invention. In addition, stress-related fusion proteins, antigenic peptides, and anti-stress-related antibodies are encompassed. The invention also provides recombinant expression vectors containing a nucleic acid molecule of the invention and cells into which the expression vectors have been introduced. Methods for producing the polypeptides and methods of use for the polypeptides of the invention are further disclosed.
Claims26
An isolated nucleic acid molecule selected from the group consisting of:
- a) a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163;
- b) a nucleic acid molecule comprising a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163;
- c) a nucleic acid molecule that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164;
- d) a nucleic acid molecule comprising a nucleotide sequence encoding a polypeptide having at least 80% amino acid sequence identity to the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164;
- and,
- e) a complement of any of (a)-(d).
An isolated polypeptide selected from the group consisting of:
- a) a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164;
- b) a polypeptide comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164, wherein said polypeptide retains activity;
- c) a polypeptide encoded by the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163;
- and,
- d) a polypeptide that is encoded by a nucleic acid molecule comprising a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163, wherein said polypeptide retains activity.
A method for detecting the presence of a polypeptide in a sample, comprising contacting the sample with a compound that selectively binds to the polypeptide and detecting the binding of the compound to the polypeptide, wherein said polypeptide is selected from the group consisting of:
- a) a polypeptide encoded by the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163;
- b) a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164;
- c) a polypeptide encoded by a nucleic acid sequence having at least 80% sequence identity to the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163, wherein said polypeptide retains activity;
- and,
- d) a polypeptide comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164, wherein said polypeptide retains activity.
A method for increasing the ability of a microorganism to survive stressful conditions, comprising introducing into said microorganism a nucleic acid molecule comprising at least one nucleotide sequence selected from the group consisting of:
- a) the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163;
- b) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164;
- c) a nucleotide sequence that is at least 80% identical to the sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163, wherein said nucleotide sequence encodes a polypeptide that retains activity;
- and,
- d) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164, wherein said polypeptide retains activity.
A method for enhancing the ability of a microorganism to survive passage through the gastrointestinal tract, comprising introducing into said microorganism a nucleic acid molecule comprising at least one nucleotide sequence selected from the group consisting of:
- a) the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163;
- b) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164;
- c) a nucleotide sequence that is at least 80% identical to the sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163, wherein said nucleotide sequence encodes a polypeptide that retains activity;
- and,
- d) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164, wherein said polypeptide retains activity.
A method for increasing the ability of a microorganism to survive in the presence of an antimicrobial, comprising introducing into said microorganism a nucleic acid molecule comprising at least one nucleotide sequence selected from the group consisting of:
- a) the nucleotide sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163;
- b) a nucleotide sequence encoding a polypeptide comprising the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164;
- c) a nucleotide sequence that is at least 80% identical to the sequence of SEQ ID NO:1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 49, 51 53, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 113, 115, 117, 119, 121, 123, 125, 127, 129, 143, 145, 147, 149, 151, 153, 155, 157, 159, 161 or 163, wherein said nucleotide sequence encodes a polypeptide that retains activity;
- and,
- d) a nucleotide sequence encoding a polypeptide comprising an amino acid sequence having at least 80% sequence identity to the amino acid sequence of SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 114, 116, 118, 120, 122, 124, 126, 128, 130, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162 or 164, wherein said polypeptide retains activity.
A method for enabling an organism to respond to an environmental stimuli, comprising introducing into said organism a vector comprising at least one nucleotide sequence encoding a histidine kinase selected from the group consisting of SEQ ID NOS:4, 8, 14, 16, 20, 24, 30, 34, and 36, and a response regulator selected from the group consisting of SEQ ID NOS:2, 6, 10, 12, 18, 22, 26, 28, 32, and 38.
A Lactobacillus acidophilus cell with an increased ability to survive stressful conditions compared to a wild-type Lactobacillus acidophilus cell, wherein said increased ability to survive stressful conditions is the result of overexpression of a nucleic acid molecule encoding an amino acid sequence as set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, or 102.
A Lactobacillus acidophilus cell with an increased ability to survive osmotic stress compared to a wild-type Lactobacillus acidophilus cell, wherein said increased ability to survive osmotic stress is the result of overexpression of a nucleic acid molecule encoding an amino acid sequence as set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, or 102.
A Lactobacillus acidophilus cell with an increased ability to survive oxidative stress compared to a wild-type Lactobacillus acidophilus cell, wherein said increased ability to survive oxidative stress is the result of overexpression of a nucleic acid molecule encoding an amino acid sequence as set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, or 102.
A Lactobacillus acidophilus cell with an increased ability to survive starvation conditions compared to a wild-type Lactobacillus acidophilus cell, wherein said increased ability to survive starvation conditions is the result of overexpression of a nucleic acid molecule encoding an amino acid sequence as set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, or 102.
A Lactobacillus acidophilus cell with an increased ability to survive in the presence of antimicrobials compared to a wild-type Lactobacillus acidophilus cell, wherein said increased ability to survive in the presence of antimicrobials is the result of overexpression of a nucleic acid molecule encoding an amino acid sequence as set forth in SEQ ID NO:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 50, 52, 54, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, 94, 96, 98, 100, or 102.
Dependencies are inferred from the claim text.