DK2814954T3 - endonucleases - Google Patents

Info

Publication number

DK2814954T3

DK2814954T3 DK13705249.4T DK13705249T DK2814954T3 DK 2814954 T3 DK2814954 T3 DK 2814954T3 DK 13705249 T DK13705249 T DK 13705249T DK 2814954 T3 DK2814954 T3 DK 2814954T3

Authority

DK

Denmark

Prior art keywords

gin

gly

ala

arg

lys

Prior art date

2012-02-17

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• 108010042407 Endonucleases Proteins 0.000 title claims description 115
• 102000004533 Endonucleases Human genes 0.000 title claims description 31
• FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 70
• 108020004414 DNA Proteins 0.000 claims description 64
• 108010008532 Deoxyribonuclease I Proteins 0.000 claims description 59
• 102000007260 Deoxyribonuclease I Human genes 0.000 claims description 59
• 102000004190 Enzymes Human genes 0.000 claims description 59
• 108090000790 Enzymes Proteins 0.000 claims description 59
• 238000000034 method Methods 0.000 claims description 57
• 230000000694 effects Effects 0.000 claims description 54
• PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 claims description 44
• 150000007523 nucleic acids Chemical class 0.000 claims description 42
• 108090000623 proteins and genes Proteins 0.000 claims description 42
• VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 claims description 41
• 102000039446 nucleic acids Human genes 0.000 claims description 41
• 108020004707 nucleic acids Proteins 0.000 claims description 41
• 239000000654 additive Substances 0.000 claims description 40
• 102000004169 proteins and genes Human genes 0.000 claims description 39
• 239000012634 fragment Substances 0.000 claims description 38
• 235000018102 proteins Nutrition 0.000 claims description 36
• 230000000996 additive effect Effects 0.000 claims description 35
• 239000011780 sodium chloride Substances 0.000 claims description 35
• 239000000203 mixture Substances 0.000 claims description 34
• WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 23
• 102000040430 polynucleotide Human genes 0.000 claims description 22
• 108091033319 polynucleotide Proteins 0.000 claims description 22
• 239000002157 polynucleotide Substances 0.000 claims description 22
• 239000003153 chemical reaction reagent Substances 0.000 claims description 18
• 150000003839 salts Chemical class 0.000 claims description 18
• 239000000243 solution Substances 0.000 claims description 17
• 238000000746 purification Methods 0.000 claims description 15
• 241000122170 Aliivibrio salmonicida Species 0.000 claims description 13
• WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 12
• DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 claims description 11
• 241000607598 Vibrio Species 0.000 claims description 10
• 125000000539 amino acid group Chemical group 0.000 claims description 10
• 238000003556 assay Methods 0.000 claims description 10
• 230000003197 catalytic effect Effects 0.000 claims description 9
• 102000052510 DNA-Binding Proteins Human genes 0.000 claims description 8
• KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 7
• 239000003638 chemical reducing agent Substances 0.000 claims description 7
• 235000013922 glutamic acid Nutrition 0.000 claims description 7
• 239000004220 glutamic acid Substances 0.000 claims description 7
• 239000013592 cell lysate Substances 0.000 claims description 6
• 101710096438 DNA-binding protein Proteins 0.000 claims description 4
• 238000001712 DNA sequencing Methods 0.000 claims description 3
• 239000002738 chelating agent Substances 0.000 claims description 3
• 238000002955 isolation Methods 0.000 claims description 3
• 229910021645 metal ion Inorganic materials 0.000 claims description 3
• 230000008569 process Effects 0.000 claims description 3
• JPSHPWJJSVEEAX-NFJMKROFSA-N (2s)-2-amino-4-fluoropentanedioic acid Chemical compound OC(=O)[C@@H](N)CC(F)C(O)=O JPSHPWJJSVEEAX-NFJMKROFSA-N 0.000 claims description 2
• OYIFNHCXNCRBQI-SCSAIBSYSA-N D-2-aminoadipic acid Chemical compound OC(=O)[C@H](N)CCCC(O)=O OYIFNHCXNCRBQI-SCSAIBSYSA-N 0.000 claims description 2
• 108010021625 Immunoglobulin Fragments Proteins 0.000 claims description 2
• 102000008394 Immunoglobulin Fragments Human genes 0.000 claims description 2
• CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 2
• GHAZCVNUKKZTLG-UHFFFAOYSA-N N-ethyl-succinimide Natural products CCN1C(=O)CCC1=O GHAZCVNUKKZTLG-UHFFFAOYSA-N 0.000 claims description 2
• HDFGOPSGAURCEO-UHFFFAOYSA-N N-ethylmaleimide Chemical compound CCN1C(=O)C=CC1=O HDFGOPSGAURCEO-UHFFFAOYSA-N 0.000 claims description 2
• 238000003559 RNA-seq method Methods 0.000 claims description 2
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• 235000003704 aspartic acid Nutrition 0.000 claims description 2
• OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 2
• 210000001124 body fluid Anatomy 0.000 claims description 2
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• UHBYWPGGCSDKFX-UHFFFAOYSA-N carboxyglutamic acid Chemical compound OC(=O)C(N)CC(C(O)=O)C(O)=O UHBYWPGGCSDKFX-UHFFFAOYSA-N 0.000 claims description 2
• 125000000291 glutamic acid group Chemical group N[C@@H](CCC(O)=O)C(=O)* 0.000 claims description 2
• 239000000126 substance Substances 0.000 claims description 2
• 230000000415 inactivating effect Effects 0.000 claims 2
• OGMADIBCHLQMIP-UHFFFAOYSA-N 2-aminoethanethiol;hydron;chloride Chemical compound Cl.NCCS OGMADIBCHLQMIP-UHFFFAOYSA-N 0.000 claims 1
• 238000003776 cleavage reaction Methods 0.000 claims 1
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• 239000000523 sample Substances 0.000 description 33
• SSZGOKWBHLOCHK-DCAQKATOSA-N Arg-Lys-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCN=C(N)N SSZGOKWBHLOCHK-DCAQKATOSA-N 0.000 description 29
• SYELGNBERZZXAG-JQWIXIFHSA-N Cys-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CS)N)C(O)=O)=CNC2=C1 SYELGNBERZZXAG-JQWIXIFHSA-N 0.000 description 29
• 108010051242 phenylalanylserine Proteins 0.000 description 28
• XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 27
• TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 26
• XEXSSIBQYNKFBX-KBPBESRZSA-N Phe-Gly-His Chemical compound C([C@H](N)C(=O)NCC(=O)N[C@@H](CC=1N=CNC=1)C(O)=O)C1=CC=CC=C1 XEXSSIBQYNKFBX-KBPBESRZSA-N 0.000 description 26
• 238000006243 chemical reaction Methods 0.000 description 26
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• DUYYPIRFTLOAJQ-YUMQZZPRSA-N Gly-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)CN DUYYPIRFTLOAJQ-YUMQZZPRSA-N 0.000 description 25
• JSNNHGHYGYMVCK-XVKPBYJWSA-N Gly-Glu-Val Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O JSNNHGHYGYMVCK-XVKPBYJWSA-N 0.000 description 25
• IICZCLFBILYRCU-WHFBIAKZSA-N Asn-Gly-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IICZCLFBILYRCU-WHFBIAKZSA-N 0.000 description 24
• 238000003753 real-time PCR Methods 0.000 description 24
• JAMAWBXXKFGFGX-KZVJFYERSA-N Ala-Arg-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O JAMAWBXXKFGFGX-KZVJFYERSA-N 0.000 description 22
• MHNHRNHJMXAVHZ-AAEUAGOBSA-N Trp-Asn-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)O)N MHNHRNHJMXAVHZ-AAEUAGOBSA-N 0.000 description 22
• 150000001413 amino acids Chemical group 0.000 description 22
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• URDUGPGPLNXXES-WHFBIAKZSA-N Cys-Gly-Cys Chemical compound SC[C@H](N)C(=O)NCC(=O)N[C@@H](CS)C(O)=O URDUGPGPLNXXES-WHFBIAKZSA-N 0.000 description 20
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• ONPFOYPPPOHMNH-UVBJJODRSA-N Pro-Ala-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@@H]3CCCN3 ONPFOYPPPOHMNH-UVBJJODRSA-N 0.000 description 20
• 108010064235 lysylglycine Proteins 0.000 description 20
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• 108010069205 aspartyl-phenylalanine Proteins 0.000 description 18
• VJVQKGYHIZPSNS-FXQIFTODSA-N Ala-Ser-Arg Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N VJVQKGYHIZPSNS-FXQIFTODSA-N 0.000 description 17
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• BPCLDCNZBUYGOD-BPUTZDHNSA-N Glu-Trp-Glu Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 BPCLDCNZBUYGOD-BPUTZDHNSA-N 0.000 description 17
• VEPBEGNDJYANCF-QWRGUYRKSA-N Gly-Lys-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCCN VEPBEGNDJYANCF-QWRGUYRKSA-N 0.000 description 17
• 108010076504 Protein Sorting Signals Proteins 0.000 description 17
• HPYDSVWYXXKHRD-VIFPVBQESA-N Tyr-Gly Chemical compound [O-]C(=O)CNC(=O)[C@@H]([NH3+])CC1=CC=C(O)C=C1 HPYDSVWYXXKHRD-VIFPVBQESA-N 0.000 description 17
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