AU2006207308A1 - Polypeptide mutagenesis method - Google Patents
Polypeptide mutagenesis method Download PDFInfo
- Publication number
- AU2006207308A1 AU2006207308A1 AU2006207308A AU2006207308A AU2006207308A1 AU 2006207308 A1 AU2006207308 A1 AU 2006207308A1 AU 2006207308 A AU2006207308 A AU 2006207308A AU 2006207308 A AU2006207308 A AU 2006207308A AU 2006207308 A1 AU2006207308 A1 AU 2006207308A1
- Authority
- AU
- Australia
- Prior art keywords
- transposon
- dna
- sequence
- restriction enzyme
- amino acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 238000000034 method Methods 0.000 title claims abstract description 119
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 35
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 35
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 35
- 238000002703 mutagenesis Methods 0.000 title description 19
- 231100000350 mutagenesis Toxicity 0.000 title description 19
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract description 102
- 108091008146 restriction endonucleases Proteins 0.000 claims abstract description 89
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 41
- 230000007018 DNA scission Effects 0.000 claims abstract description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 175
- 108020004414 DNA Proteins 0.000 claims description 129
- 238000003780 insertion Methods 0.000 claims description 117
- 230000037431 insertion Effects 0.000 claims description 116
- 125000003729 nucleotide group Chemical group 0.000 claims description 107
- 239000002773 nucleotide Substances 0.000 claims description 105
- 102000004169 proteins and genes Human genes 0.000 claims description 82
- 239000013612 plasmid Substances 0.000 claims description 74
- 238000012217 deletion Methods 0.000 claims description 47
- 230000037430 deletion Effects 0.000 claims description 45
- 238000006243 chemical reaction Methods 0.000 claims description 41
- 230000035772 mutation Effects 0.000 claims description 39
- 150000001413 amino acids Chemical class 0.000 claims description 38
- 230000017105 transposition Effects 0.000 claims description 37
- 238000006467 substitution reaction Methods 0.000 claims description 35
- 102000004190 Enzymes Human genes 0.000 claims description 28
- 108090000790 Enzymes Proteins 0.000 claims description 28
- 230000029087 digestion Effects 0.000 claims description 28
- 239000012634 fragment Substances 0.000 claims description 26
- 230000000694 effects Effects 0.000 claims description 23
- 108010020764 Transposases Proteins 0.000 claims description 21
- 102000008579 Transposases Human genes 0.000 claims description 21
- 108020004705 Codon Proteins 0.000 claims description 20
- 108700026244 Open Reading Frames Proteins 0.000 claims description 16
- 238000003776 cleavage reaction Methods 0.000 claims description 15
- 230000007017 scission Effects 0.000 claims description 15
- 108091034117 Oligonucleotide Proteins 0.000 claims description 11
- 230000027455 binding Effects 0.000 claims description 10
- 108020001580 protein domains Proteins 0.000 claims description 10
- 230000009466 transformation Effects 0.000 claims description 10
- 238000000926 separation method Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 6
- 238000012216 screening Methods 0.000 claims description 6
- 238000012163 sequencing technique Methods 0.000 claims description 6
- 239000000872 buffer Substances 0.000 claims description 5
- 230000003281 allosteric effect Effects 0.000 claims description 4
- 108091081024 Start codon Proteins 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 2
- 210000004027 cell Anatomy 0.000 description 59
- 101150049515 bla gene Proteins 0.000 description 48
- 241000588724 Escherichia coli Species 0.000 description 35
- 101000740462 Escherichia coli Beta-lactamase TEM Proteins 0.000 description 31
- 239000013615 primer Substances 0.000 description 30
- 229960000723 ampicillin Drugs 0.000 description 29
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 29
- 229960005091 chloramphenicol Drugs 0.000 description 23
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 23
- 239000000203 mixture Substances 0.000 description 20
- 239000003550 marker Substances 0.000 description 16
- 239000006142 Luria-Bertani Agar Substances 0.000 description 14
- 101150011679 cybC gene Proteins 0.000 description 14
- 238000005516 engineering process Methods 0.000 description 14
- 238000000246 agarose gel electrophoresis Methods 0.000 description 13
- 230000007246 mechanism Effects 0.000 description 13
- 238000004520 electroporation Methods 0.000 description 12
- 238000004458 analytical method Methods 0.000 description 11
- 101100494389 Yersinia pestis cybC1 gene Proteins 0.000 description 10
- 101100494390 Yersinia pestis cybC2 gene Proteins 0.000 description 10
- 102000012410 DNA Ligases Human genes 0.000 description 9
- 108010061982 DNA Ligases Proteins 0.000 description 9
- 230000006870 function Effects 0.000 description 9
- 108010006785 Taq Polymerase Proteins 0.000 description 8
- 238000000338 in vitro Methods 0.000 description 8
- 239000011541 reaction mixture Substances 0.000 description 8
- 230000008901 benefit Effects 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 230000012010 growth Effects 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 238000013459 approach Methods 0.000 description 6
- 230000002068 genetic effect Effects 0.000 description 6
- 230000008439 repair process Effects 0.000 description 6
- 230000002441 reversible effect Effects 0.000 description 6
- 238000010367 cloning Methods 0.000 description 5
- 238000010276 construction Methods 0.000 description 5
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- 230000010076 replication Effects 0.000 description 5
- 239000000758 substrate Substances 0.000 description 5
- 102100031780 Endonuclease Human genes 0.000 description 4
- 108010042407 Endonucleases Proteins 0.000 description 4
- 102220474982 Histidine-tRNA ligase, cytoplasmic_R83A_mutation Human genes 0.000 description 4
- 238000012300 Sequence Analysis Methods 0.000 description 4
- 101000623262 Trypanosoma brucei brucei Uncharacterized 22 kDa protein in aldolase locus Proteins 0.000 description 4
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 4
- 108020004707 nucleic acids Proteins 0.000 description 4
- 102000039446 nucleic acids Human genes 0.000 description 4
- 150000007523 nucleic acids Chemical class 0.000 description 4
- SCVFZCLFOSHCOH-UHFFFAOYSA-M potassium acetate Chemical compound [K+].CC([O-])=O SCVFZCLFOSHCOH-UHFFFAOYSA-M 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 239000011535 reaction buffer Substances 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 239000006137 Luria-Bertani broth Substances 0.000 description 3
- 108091006629 SLC13A2 Proteins 0.000 description 3
- 238000007792 addition Methods 0.000 description 3
- 239000003782 beta lactam antibiotic agent Substances 0.000 description 3
- 230000033228 biological regulation Effects 0.000 description 3
- 229940098773 bovine serum albumin Drugs 0.000 description 3
- 210000004899 c-terminal region Anatomy 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 230000002779 inactivation Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000013519 translation Methods 0.000 description 3
- 102000053602 DNA Human genes 0.000 description 2
- 238000001712 DNA sequencing Methods 0.000 description 2
- IEGFSKKANYKBDU-QWHCGFSZSA-N Gly-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)CN)C(=O)O IEGFSKKANYKBDU-QWHCGFSZSA-N 0.000 description 2
- 102100024319 Intestinal-type alkaline phosphatase Human genes 0.000 description 2
- 101710184243 Intestinal-type alkaline phosphatase Proteins 0.000 description 2
- 206010034133 Pathogen resistance Diseases 0.000 description 2
- 101150090155 R gene Proteins 0.000 description 2
- 229920004890 Triton X-100 Polymers 0.000 description 2
- 239000013504 Triton X-100 Substances 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 239000011543 agarose gel Substances 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 244000309466 calf Species 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 230000000295 complement effect Effects 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 230000037433 frameshift Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000011835 investigation Methods 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000003446 ligand Substances 0.000 description 2
- UEGPKNKPLBYCNK-UHFFFAOYSA-L magnesium acetate Chemical compound [Mg+2].CC([O-])=O.CC([O-])=O UEGPKNKPLBYCNK-UHFFFAOYSA-L 0.000 description 2
- 229940069446 magnesium acetate Drugs 0.000 description 2
- 235000011285 magnesium acetate Nutrition 0.000 description 2
- 239000011654 magnesium acetate Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000005257 nucleotidylation Effects 0.000 description 2
- 235000011056 potassium acetate Nutrition 0.000 description 2
- 230000037432 silent mutation Effects 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- ATHGHQPFGPMSJY-UHFFFAOYSA-N spermidine Chemical compound NCCCCNCCCN ATHGHQPFGPMSJY-UHFFFAOYSA-N 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- PIEPQKCYPFFYMG-UHFFFAOYSA-N tris acetate Chemical compound CC(O)=O.OCC(N)(CO)CO PIEPQKCYPFFYMG-UHFFFAOYSA-N 0.000 description 2
- 108700026215 vpr Genes Proteins 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- DBGSRZSKGVSXRK-UHFFFAOYSA-N 1-[2-[5-[2-(2,3-dihydro-1H-inden-2-ylamino)pyrimidin-5-yl]-1,3,4-oxadiazol-2-yl]acetyl]-3,6-dihydro-2H-pyridine-4-carboxylic acid Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C1=NN=C(O1)CC(=O)N1CCC(=CC1)C(=O)O DBGSRZSKGVSXRK-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 101150098072 20 gene Proteins 0.000 description 1
- 101150110188 30 gene Proteins 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 108010089921 CTCGAG-specific type II deoxyribonucleases Proteins 0.000 description 1
- 102000005701 Calcium-Binding Proteins Human genes 0.000 description 1
- 108010045403 Calcium-Binding Proteins Proteins 0.000 description 1
- 101710205889 Cytochrome b562 Proteins 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 208000034454 F12-related hereditary angioedema with normal C1Inh Diseases 0.000 description 1
- 108010036162 GATC-specific type II deoxyribonucleases Proteins 0.000 description 1
- JBCLFWXMTIKCCB-VIFPVBQESA-N Gly-Phe Chemical compound NCC(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-VIFPVBQESA-N 0.000 description 1
- BCCRXDTUTZHDEU-VKHMYHEASA-N Gly-Ser Chemical compound NCC(=O)N[C@@H](CO)C(O)=O BCCRXDTUTZHDEU-VKHMYHEASA-N 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000012330 Integrases Human genes 0.000 description 1
- 108010061833 Integrases Proteins 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- 108010021757 Polynucleotide 5'-Hydroxyl-Kinase Proteins 0.000 description 1
- 102000008422 Polynucleotide 5'-hydroxyl-kinase Human genes 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- 108090000787 Subtilisin Proteins 0.000 description 1
- 108010012306 Tn5 transposase Proteins 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000036755 cellular response Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- SUYVUBYJARFZHO-RRKCRQDMSA-N dATP Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-RRKCRQDMSA-N 0.000 description 1
- SUYVUBYJARFZHO-UHFFFAOYSA-N dATP Natural products C1=NC=2C(N)=NC=NC=2N1C1CC(O)C(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 SUYVUBYJARFZHO-UHFFFAOYSA-N 0.000 description 1
- RGWHQCVHVJXOKC-SHYZEUOFSA-J dCTP(4-) Chemical compound O=C1N=C(N)C=CN1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)C1 RGWHQCVHVJXOKC-SHYZEUOFSA-J 0.000 description 1
- HAAZLUGHYHWQIW-KVQBGUIXSA-N dGTP Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@H]1C[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)O1 HAAZLUGHYHWQIW-KVQBGUIXSA-N 0.000 description 1
- NHVNXKFIZYSCEB-XLPZGREQSA-N dTTP Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP(O)(O)=O)[C@@H](O)C1 NHVNXKFIZYSCEB-XLPZGREQSA-N 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000002939 deleterious effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000010429 evolutionary process Effects 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000003209 gene knockout Methods 0.000 description 1
- 102000054767 gene variant Human genes 0.000 description 1
- KZNQNBZMBZJQJO-YFKPBYRVSA-N glyclproline Chemical compound NCC(=O)N1CCC[C@H]1C(O)=O KZNQNBZMBZJQJO-YFKPBYRVSA-N 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 208000016861 hereditary angioedema type 3 Diseases 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000005304 joining Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000003278 mimic effect Effects 0.000 description 1
- 108091005763 multidomain proteins Proteins 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 230000037452 priming Effects 0.000 description 1
- 230000004952 protein activity Effects 0.000 description 1
- 230000004850 protein–protein interaction Effects 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 229940063673 spermidine Drugs 0.000 description 1
- 239000012089 stop solution Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000002463 transducing effect Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 241001515965 unidentified phage Species 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/1034—Isolating an individual clone by screening libraries
- C12N15/1093—General methods of preparing gene libraries, not provided for in other subgroups
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
- C12N15/102—Mutagenizing nucleic acids
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Crystallography & Structural Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Plant Pathology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Computational Biology (AREA)
- Enzymes And Modification Thereof (AREA)
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Laminated Bodies (AREA)
- Wrappers (AREA)
- Polyoxymethylene Polymers And Polymers With Carbon-To-Carbon Bonds (AREA)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GBGB0501189.5A GB0501189D0 (en) | 2005-01-20 | 2005-01-20 | Polypeptide mutagenesis method |
| GB0501189.5 | 2005-01-20 | ||
| PCT/GB2006/000187 WO2006077411A1 (en) | 2005-01-20 | 2006-01-19 | Polypeptide mutagenesis method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| AU2006207308A1 true AU2006207308A1 (en) | 2006-07-27 |
Family
ID=34259403
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| AU2006207308A Abandoned AU2006207308A1 (en) | 2005-01-20 | 2006-01-19 | Polypeptide mutagenesis method |
Country Status (10)
| Country | Link |
|---|---|
| US (1) | US20090053761A1 (enExample) |
| EP (1) | EP1838851B1 (enExample) |
| JP (1) | JP2008527987A (enExample) |
| AT (1) | ATE443137T1 (enExample) |
| AU (1) | AU2006207308A1 (enExample) |
| CA (1) | CA2601324A1 (enExample) |
| DE (1) | DE602006009217D1 (enExample) |
| DK (1) | DK1838851T3 (enExample) |
| GB (1) | GB0501189D0 (enExample) |
| WO (1) | WO2006077411A1 (enExample) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0515131D0 (en) * | 2005-07-22 | 2005-08-31 | Univ Aston | Oligonucleotide library encoding randomised peptides |
| US20150337290A1 (en) * | 2012-09-07 | 2015-11-26 | Alexander Gubin | Methods and products for mutating nucleotide sequences |
| LT3027771T (lt) * | 2013-07-30 | 2019-04-25 | Gen9, Inc. | Ilgos kloninės sekos verifikuotų nukleorūgščių konstruktų gamybos būdas |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4830965A (en) * | 1983-12-12 | 1989-05-16 | Canadian Patents And Development Ltd. | Transposable linkers to transfer genetic information |
| US5677170A (en) * | 1994-03-02 | 1997-10-14 | The Johns Hopkins University | In vitro transposition of artificial transposons |
| DE10109517A1 (de) * | 2001-02-28 | 2002-08-29 | Volker Sieber | Verfahren zur Herstellung von Genbibliotheken |
| US20020119573A1 (en) | 2001-02-28 | 2002-08-29 | Shaw Karen J. | Footprinting plasmid |
| JP2002253255A (ja) * | 2001-03-01 | 2002-09-10 | Japan Science & Technology Corp | ランダム挿入削除ｄｎａ変異法 |
| FI20020746A7 (fi) * | 2002-04-18 | 2003-10-19 | Finnzymes Oy | Menetelmä ja materiaalit polypeptidien deleetiojohdannaisten tuottamiseksi |
| KR100391884B1 (en) * | 2002-07-04 | 2003-09-06 | Amicogen Co Ltd | Random codon based mutagenesis using transposon |
-
2005
- 2005-01-20 GB GBGB0501189.5A patent/GB0501189D0/en not_active Ceased
-
2006
- 2006-01-19 EP EP06703246A patent/EP1838851B1/en not_active Not-in-force
- 2006-01-19 JP JP2007551737A patent/JP2008527987A/ja active Pending
- 2006-01-19 US US11/795,732 patent/US20090053761A1/en not_active Abandoned
- 2006-01-19 AT AT06703246T patent/ATE443137T1/de not_active IP Right Cessation
- 2006-01-19 AU AU2006207308A patent/AU2006207308A1/en not_active Abandoned
- 2006-01-19 CA CA002601324A patent/CA2601324A1/en not_active Abandoned
- 2006-01-19 WO PCT/GB2006/000187 patent/WO2006077411A1/en not_active Ceased
- 2006-01-19 DE DE602006009217T patent/DE602006009217D1/de active Active
- 2006-01-19 DK DK06703246.6T patent/DK1838851T3/da active
Also Published As
| Publication number | Publication date |
|---|---|
| EP1838851A1 (en) | 2007-10-03 |
| US20090053761A1 (en) | 2009-02-26 |
| ATE443137T1 (de) | 2009-10-15 |
| DK1838851T3 (da) | 2010-01-25 |
| CA2601324A1 (en) | 2006-07-27 |
| JP2008527987A (ja) | 2008-07-31 |
| EP1838851B1 (en) | 2009-09-16 |
| DE602006009217D1 (en) | 2009-10-29 |
| WO2006077411A1 (en) | 2006-07-27 |
| GB0501189D0 (en) | 2005-03-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11098326B2 (en) | Using RNA-guided FokI nucleases (RFNs) to increase specificity for RNA-guided genome editing | |
| JP7187435B2 (ja) | ゲノム編集 | |
| JP4020429B2 (ja) | 操作された組換え部位を使用する組換えクローニング | |
| US20030068799A1 (en) | Recombinational cloning using engineered recombination sites | |
| WO2011053957A9 (en) | Compositions and methods for the regulation of multiple genes of interest in a cell | |
| WO2014204578A1 (en) | Using rna-guided foki nucleases (rfns) to increase specificity for rna-guided genome editing | |
| WO1996040724A9 (en) | Recombinational cloning using engineered recombination sites | |
| WO2002044409A2 (en) | Substrate linked directed evolution (slide) | |
| EP1838851B1 (en) | Polypeptide mutagenesis method | |
| JP2024543216A (ja) | Ｃｒｉｓｐｒ関連トランスポザーゼおよびその使用方法 | |
| US7160702B2 (en) | Methods and nucleic acid vectors for rapid expression and screening of CDNA clones | |
| AU2002325588B2 (en) | A composition comprising nucleic acids | |
| NZ516384A (en) | Composition comprising a nucleic acid molecule | |
| AU2007242911A1 (en) | Recombinational cloning using engineered recombination sites | |
| AU1006201A (en) | Recombinational cloning using engineered recombination sites | |
| HK40002080B (en) | Genome editing | |
| HK40002080A (en) | Genome editing | |
| Rasila | Functional Mapping of Mu Transposition Machinery: MuA Protein Modification and Engineering for Hyperactivity | |
| HK1019068B (en) | Recombinational cloning in vitro using engineered recombination sites |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MK4 | Application lapsed section 142(2)(d) - no continuation fee paid for the application |