Method to isolate mutants and to clone the complementing gene
The subject invention lies in the field of microorganism mutation and selection of the mutants. In particular, the invention is directed at obtaining metabolic mutants in a simple, direct and specific manner. In a preferred embodiment it is also possible to obtain desired mutants not comprising recombinant DNA, thereby facilitating incorporation thereof in products for human consumption or application, due to shorter legislative procedures. The method according to the invention involves random mutation and specific selection of the desired metabolic mutant. Knockout, mutants wherein a gene associated with metabolism is absent or inactivated and mutants with increased or decreased DNA binding capacity are also claimed.
1. An isolated nucleic acid molecule comprising nucleotides 1 to 4173 of SEQ ID NO:9.
2. An isolated nucleic acid molecule encoding amino acid residues 1 to 875 of SEQ ID NO: 11.
3. An isolated nucleic acid molecule having at least 96% identity to nucleotides 1 to 4173 of SEQ ID NO:9, wherein said nucleic acid molecule encodes a xylanolytic regulator comprising a zinc finger binding domain, wherein said xylanolytic regulator is capable of binding to one or more of target genes selected from the group consisting of xlnA, xlnB, xlnC, xlnD and axeA.
4. The isolated nucleic acid molecule of claim 3 , wherein said nucleic acid molecule has at least 97% identity to nucleotides 1 to 4173 of SEQ ID NO:9, wherein said nucleic acid molecule encodes a xylanolytic regulator comprising a zinc finger binding domain, wherein said xylanolytic regulator is capable of binding to one or more of target genes selected from the group consisting of xlnA, xlnB, xlnC, xlnD and axeA.
5. The isolated nucleic acid molecule of claim 3 , wherein said nucleic acid molecule has at least 98% identity to nucleotides 1 to 4173 of SEQ ID NO:9, wherein said nucleic acid molecule encodes a xylanolytic regulator comprising a zinc finger binding domain, wherein said xylanolytic regulator is capable of binding to one or more of target genes selected from the group consisting of xlnA, xlnB, xlnC, xlnD and axeA.
6. The isolated nucleic acid molecule of claim 3 , wherein said nucleic acid molecule has at least 99% identity to nucleotides 1 to 4173 of SEQ ID NO:9, wherein said nucleic acid molecule encodes a xylanolytic regulator comprising a zinc finger binding domain, wherein said xylanolytic regulator is capable of binding to one or more of target genes selected from the group consisting of xlnA, xlnB, xlnC, xlnD and axeA.
7. An isolated nucleic acid molecule having at least 96% identity with a nucleic acid molecule encoding amino acid residues 1 to 875 of SEQ ID NO: 11, wherein said isolated nucleic acid molecule encodes a xylanolytic regulator comprising a zinc finger binding domain, wherein said xylanolytic regulator is capable of binding to one or more of target genes selected from the group consisting of xlnA, xlnB, xlnC, xlnD and axeA.
8. The isolated nucleic acid molecule of claim 7 , wherein said isolated nucleic acid molecule has at least 97% identity to a nucleic acid molecule encoding amino acid residues 1 to 875 of SEQ ID NO: 11, wherein said nucleic acid molecule encodes a xylanolytic regulator comprising a zinc finger binding domain, wherein said xylanolytic regulator is capable of binding to one or more of target genes selected from the group consisting of xlnA, xlnB, xlnC, xlnD and axeA.
9. The isolated nucleic acid molecule of claim 7 , wherein said isolated nucleic acid molecule has at least 98% identity to a nucleic acid molecule encoding amino acid residues 1 to 875 of SEQ ID NO: 11, wherein said nucleic acid molecule encodes a xylanolytic regulator comprising a zinc finger binding domain, wherein said xylanolytic regulator is capable of binding to one or more of target genes selected from the group consisting of xlnA, xlnB, xlnC, xlnD and axeA.
10. The isolated nucleic acid molecule of claim 7 , wherein said isolated nucleic acid molecule has at least 99% identity to a nucleic acid molecule encoding amino acid residues 1 to 875 of SEQ ID NO: 11, wherein said nucleic acid molecule encodes a xylanolytic regulator comprising a zinc finger binding domain, wherein said xylanolytic regulator is capable of binding to one or more of target genes selected from the group consisting of xlnA, xlnB, xlnC, xlnD and axeA.
11. A kit for detecting and/or isolating nucleic acid sequences wherein said kit comprises two or more nucleic acid molecules identical to a portion of SEQ ID NO: 9, wherein in each of the two or more nucleic acid molecules the portion identical to SEQ ID NO: 9 is at least 60 nucleotides in length, and wherein each of the two or more nucleic acid molecules do not encode a zinc finger binding domain.