{"id":"https://openalex.org/W57726098","doi":null,"title":"Directed evolution of Thermus aquaticus DNA polymerase by compartmentalised self-replication.","display_name":"Directed evolution of Thermus aquaticus DNA polymerase by compartmentalised self-replication.","publication_year":2009,"publication_date":"2009-01-01","ids":{"openalex":"https://openalex.org/W57726098","mag":"57726098"},"language":"en","primary_location":{"id":"pmh:oai:opus.bath.ac.uk:18983","doi":null,"is_oa":false,"landing_page_url":"http://opus.bath.ac.uk/18983/1/S.Lamble_Thesis_(1).pdf","pdf_url":"http://opus.bath.ac.uk/18983/1/S.Lamble_Thesis_(1).pdf","source":{"id":"https://openalex.org/S4306401631","display_name":"The University of Bath Online Publications Store (The University of Bath)","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":"https://openalex.org/I51601045","host_organization_name":"University of Bath","host_organization_lineage":["https://openalex.org/I51601045"],"host_organization_lineage_names":[],"type":"repository"},"license":null,"license_id":null,"version":"submittedVersion","is_accepted":false,"is_published":false,"raw_source_name":"","raw_type":"Thesis"},"volume":null,"type":"dissertation","is_traditional":true,"indexed_in":[],"open_access":{"is_oa":false,"oa_status":"closed","oa_url":null,"any_repository_has_fulltext":false},"authorships":[{"author_position":"first","author":{"id":null,"display_name":"Sarah Lamble","orcid":"https://orcid.org/0000-0003-3443-1004","observed_orcids":["https://orcid.org/0000-0003-3443-1004"]},"institutions":[],"countries":[],"is_corresponding":true,"raw_author_name":"Sarah Lamble","raw_affiliation_strings":["Department of Biology & Biochemistry"],"raw_orcid":"https://orcid.org/0000-0003-3443-1004","affiliations":[{"raw_affiliation_string":"Department of Biology & Biochemistry","institution_ids":[]}]}],"institutions":[],"countries_distinct_count":0,"institutions_distinct_count":0,"corresponding_author_ids":[],"corresponding_institution_ids":[],"apc_list":null,"apc_paid":null,"fwci":null,"has_fulltext":false,"cited_by_count":0,"citation_normalized_percentile":null,"cited_by_percentile_year":null,"biblio":{"volume":null,"issue":null,"first_page":null,"last_page":null},"is_retracted":false,"is_paratext":false,"is_xpac":false,"primary_topic":{"id":"https://openalex.org/T10404","display_name":"Enzyme Catalysis and Immobilization","score":0.8036999702453613,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},"topics":[{"id":"https://openalex.org/T10404","display_name":"Enzyme Catalysis and Immobilization","score":0.8036999702453613,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T11970","display_name":"Molecular Biology Techniques and Applications","score":0.07349999994039536,"subfield":{"id":"https://openalex.org/subfields/1312","display_name":"Molecular Biology"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}},{"id":"https://openalex.org/T11764","display_name":"Evolution and Genetic Dynamics","score":0.028200000524520874,"subfield":{"id":"https://openalex.org/subfields/1311","display_name":"Genetics"},"field":{"id":"https://openalex.org/fields/13","display_name":"Biochemistry, Genetics and Molecular Biology"},"domain":{"id":"https://openalex.org/domains/1","display_name":"Life Sciences"}}],"keywords":[{"id":"https://openalex.org/keywords/directed-evolution","display_name":"directed evolution","score":0.906000018119812},{"id":"https://openalex.org/keywords/pcr-inhibitors","display_name":"PCR inhibitors","score":0.8619999885559082},{"id":"https://openalex.org/keywords/heparin-resistance","display_name":"heparin resistance","score":0.9769999980926514},{"id":"https://openalex.org/keywords/error-prone-pcr","display_name":"error-prone PCR","score":0.7070000171661377},{"id":"https://openalex.org/keywords/thermostable-enzymes","display_name":"thermostable enzymes","score":0.5419999957084656},{"id":"https://openalex.org/keywords/pcr-diagnosis","display_name":"PCR diagnosis","score":0.3580000102519989},{"id":"https://openalex.org/keywords/magnesium-chelation","display_name":"magnesium chelation","score":0.6000000238418579},{"id":"https://openalex.org/keywords/polymerase-fidelity","display_name":"polymerase fidelity","score":0.4860000014305115},{"id":"https://openalex.org/keywords/processivity","display_name":"processivity","score":0.5400000214576721},{"id":"https://openalex.org/keywords/thermal-stability","display_name":"thermal stability","score":0.5389999747276306},{"id":"https://openalex.org/keywords/mutant-library-screening","display_name":"mutant library screening","score":0.3149999976158142}],"concepts":[{"id":"https://openalex.org/C2781440490","wikidata":"https://www.wikidata.org/wiki/Q137165","display_name":"Thermus aquaticus","level":4,"score":0.9388939738273621},{"id":"https://openalex.org/C2779186596","wikidata":"https://www.wikidata.org/wiki/Q412581","display_name":"Taq polymerase","level":5,"score":0.839350700378418},{"id":"https://openalex.org/C82381507","wikidata":"https://www.wikidata.org/wiki/Q416878","display_name":"Polymerase","level":3,"score":0.7080883979797363},{"id":"https://openalex.org/C112456955","wikidata":"https://www.wikidata.org/wiki/Q4118048","display_name":"Hot start PCR","level":5,"score":0.6711479425430298},{"id":"https://openalex.org/C2756471","wikidata":"https://www.wikidata.org/wiki/Q206286","display_name":"DNA polymerase","level":3,"score":0.6494019031524658},{"id":"https://openalex.org/C9418097","wikidata":"https://www.wikidata.org/wiki/Q440681","display_name":"Directed evolution","level":4,"score":0.6226834058761597},{"id":"https://openalex.org/C186008079","wikidata":"https://www.wikidata.org/wiki/Q7226557","display_name":"Polymerase chain reaction optimization","level":5,"score":0.6088495850563049},{"id":"https://openalex.org/C86803240","wikidata":"https://www.wikidata.org/wiki/Q420","display_name":"Biology","level":0,"score":0.5697778463363647},{"id":"https://openalex.org/C49105822","wikidata":"https://www.wikidata.org/wiki/Q176996","display_name":"Polymerase chain reaction","level":3,"score":0.5439621210098267},{"id":"https://openalex.org/C156385826","wikidata":"https://www.wikidata.org/wiki/Q1341059","display_name":"Multiple displacement amplification","level":5,"score":0.5240005254745483},{"id":"https://openalex.org/C131140672","wikidata":"https://www.wikidata.org/wiki/Q2031164","display_name":"Processivity","level":4,"score":0.4792768657207489},{"id":"https://openalex.org/C2779454882","wikidata":"https://www.wikidata.org/wiki/Q5205776","display_name":"DNA shuffling","level":5,"score":0.46912074089050293},{"id":"https://openalex.org/C153911025","wikidata":"https://www.wikidata.org/wiki/Q7202","display_name":"Molecular biology","level":1,"score":0.44873058795928955},{"id":"https://openalex.org/C88354546","wikidata":"https://www.wikidata.org/wiki/Q3502973","display_name":"DNA polymerase II","level":5,"score":0.4460391700267792},{"id":"https://openalex.org/C70721500","wikidata":"https://www.wikidata.org/wiki/Q177005","display_name":"Computational biology","level":1,"score":0.3556050956249237},{"id":"https://openalex.org/C54355233","wikidata":"https://www.wikidata.org/wiki/Q7162","display_name":"Genetics","level":1,"score":0.31187164783477783},{"id":"https://openalex.org/C143065580","wikidata":"https://www.wikidata.org/wiki/Q3285695","display_name":"Mutant","level":3,"score":0.30732351541519165},{"id":"https://openalex.org/C552990157","wikidata":"https://www.wikidata.org/wiki/Q7430","display_name":"DNA","level":2,"score":0.28910350799560547},{"id":"https://openalex.org/C104317684","wikidata":"https://www.wikidata.org/wiki/Q7187","display_name":"Gene","level":2,"score":0.2260001301765442},{"id":"https://openalex.org/C78063203","wikidata":"https://www.wikidata.org/wiki/Q3062596","display_name":"DNA extraction","level":4,"score":0.1413024663925171},{"id":"https://openalex.org/C157431215","wikidata":"https://www.wikidata.org/wiki/Q856465","display_name":"Nested polymerase chain reaction","level":4,"score":0.12273415923118591},{"id":"https://openalex.org/C156719811","wikidata":"https://www.wikidata.org/wiki/Q28774","display_name":"Reverse transcriptase","level":4,"score":0.09741654992103577}],"mesh":[],"locations_count":3,"locations":[{"id":"pmh:oai:opus.bath.ac.uk:18983","doi":null,"is_oa":false,"landing_page_url":"http://opus.bath.ac.uk/18983/1/S.Lamble_Thesis_(1).pdf","pdf_url":"http://opus.bath.ac.uk/18983/1/S.Lamble_Thesis_(1).pdf","source":{"id":"https://openalex.org/S4306401631","display_name":"The University of Bath Online Publications Store (The University of Bath)","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":"https://openalex.org/I51601045","host_organization_name":"University of Bath","host_organization_lineage":["https://openalex.org/I51601045"],"host_organization_lineage_names":[],"type":"repository"},"license":null,"license_id":null,"version":"submittedVersion","is_accepted":false,"is_published":false,"raw_source_name":"","raw_type":"Thesis"},{"id":"pmh:hdl:10068/962837","doi":null,"is_oa":false,"landing_page_url":"http://hdl.handle.net/10068/962837","pdf_url":null,"source":{"id":"https://openalex.org/S4377196900","display_name":"OpenGrey (Institut de l'Information Scientifique et Technique)","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":"https://openalex.org/I1295345811","host_organization_name":"Institut de l'Information Scientifique et Technique","host_organization_lineage":["https://openalex.org/I1295345811"],"host_organization_lineage_names":[],"type":"repository"},"license":null,"license_id":null,"version":"submittedVersion","is_accepted":false,"is_published":false,"raw_source_name":"","raw_type":"U - Thesis"},{"id":"mag:57726098","doi":null,"is_oa":false,"landing_page_url":"http://europepmc.org/theses/ETH/507743","pdf_url":null,"source":{"id":"https://openalex.org/S4306400806","display_name":"Europe PMC (PubMed Central)","issn_l":null,"issn":null,"is_oa":false,"is_in_doaj":false,"is_core":false,"listed_in":[],"host_organization":"https://openalex.org/I1303153112","host_organization_name":"European Bioinformatics Institute","host_organization_lineage":["https://openalex.org/I1303153112"],"host_organization_lineage_names":[],"type":"repository"},"license":null,"license_id":null,"version":null,"is_accepted":false,"is_published":false,"raw_source_name":null,"raw_type":null}],"best_oa_location":null,"sustainable_development_goals":[],"study_designs":[],"awards":[],"funders":[],"has_content":{"grobid_xml":false,"pdf":false},"content_urls":null,"referenced_works_count":0,"referenced_works":[],"related_works":["https://openalex.org/W2791749652","https://openalex.org/W4413803640","https://openalex.org/W2055528600","https://openalex.org/W201641443","https://openalex.org/W2137523339","https://openalex.org/W2246517747","https://openalex.org/W7096537469","https://openalex.org/W1972221771","https://openalex.org/W1994092462","https://openalex.org/W7154051843"],"abstract_inverted_index":{"The":[0,57,136,298,308,347,375,405],"thermophilic":[1],"enzyme,":[2],"Thermus":[3],"aquaticus":[4],"(Taq)":[5],"DNA":[6,21,30,191],"polymerase,":[7],"is":[8,32,65,144,150,204,443,489],"an":[9,41,327],"essential":[10,42,164],"tool":[11],"in":[12,22,35,44,62,117,123,241,263,295,318,360,416,429,454,501,536,550],"molecular":[13],"biology":[14],"because":[15],"of":[16,48,59,70,101,132,138,140,161,172,212,231,247,266,274,282,322,330,337,350,396,407,410,427,433,441,463,485,499,504,566,569,571],"its":[17,25,452],"ability":[18],"to":[19,54,89,94,97,105,152,184,197,268,305,363,372,401,450,495,511,527,538,559],"synthesis":[20],"vitro":[23],"and":[24,80,113,120,181,201,257,385,394,436,466,473,532,576],"inherent":[26],"thermal":[27],"stability.":[28],"Taq":[29,190,248,287,354,413,500,544],"polymerase":[31,37,192,221,245,575],"widely":[33],"used":[34,526],"the":[36,71,99,159,198,210,244,252,264,338,342,364,390,397,402,411,423,430,461,476,482,496,502,519,563,567],"chain":[38],"reaction":[39],"(PCR),":[40],"technique":[43],"a":[45,130,170,186,205,217,261,279,286,292,315,353,357,547,574],"broad":[46],"range":[47],"different":[49],"fields":[50],"from":[51,129,302,369],"academic":[52],"research":[53],"clinical":[55],"diagnostics.":[56],"use":[58],"PCR-based":[60],"tests":[61],"diagnostic":[63,553],"testing":[64],"ever":[66],"increasing;":[67],"however,":[68],"many":[69],"samples":[72],"being":[73],"tested":[74,258],"contain":[75],"substances":[76],"that":[77,278,389,481],"inhibit":[78],"PCR":[79,134,142,332,455,470,487,552,577],"prevent":[81],"target":[82],"amplification.":[83],"Many":[84],"attempts":[85],"have":[86],"been":[87],"made":[88],"engineer":[90],"polymerases":[91],"not":[92,145,514],"only":[93,243],"increase":[95,294,359],"resistance":[96,196],"overcome":[98],"problem":[100],"inhibition,":[102],"but":[103,148],"also":[104],"enhance":[106],"other":[107],"characteristics":[108],"such":[109],"as":[110,573],"fidelity,":[111],"processivity":[112],"thermostability.":[114],"Heparin,":[115],"found":[116,122,388],"blood":[118],"samples,":[119,125],"phytate,":[121,505],"faecal":[124],"are":[126],"two":[127,320,348],"examples":[128],"number":[131],"known":[133],"inhibitors.":[135],"mode":[137,440,484,568],"action":[139,428,442,570],"most":[141],"inhibitors":[143,199],"well":[146],"understood,":[147,445],"inhibition":[149,488],"thought":[151],"occur":[153],"by":[154,175,236,259,314,334,490],"enzyme":[155,520],"binding":[156],"or":[157],"through":[158],"chelation":[160],"Mg2+":[162,467],"ions":[163],"for":[165,189,209,270,422,475,542],"PCR.":[166],"In":[167],"this":[168,417,555],"project,":[169],"system":[171,254],"directed":[173],"evolution":[174],"compartmentalised":[176],"self-replication":[177],"(CSR)":[178],"was":[179,234,249,255,300,312,382,387,471],"established":[180,256],"successfully":[182],"employed":[183],"screen":[185,262],"mutant":[187,229,339,378,398],"library":[188,230,340],"variants":[193,222,233,276,414,545],"with":[194,291,310,326,356,366,546],"enhanced":[195],"heparin":[200,267,296,304,311,531],"phytate.":[202,374],"CSR":[203,253,283,323,351],"recently-established":[206],"high-throughput":[207],"method":[208],"creation":[211],"novel":[213,412,543],"polymerases,":[214],"based":[215],"on":[216,469],"feedback":[218],"loop":[219],"whereby":[220],"replicate":[223],"their":[224],"own":[225],"encoding":[226],"gene.":[227],"A":[228,457],"106":[232],"produced":[235,285],"random":[237],"mutagenesis":[238,523],"error-prone":[239,331],"PCR,":[240],"which":[242,319],"domain":[246],"mutagenised.":[250],"Firstly,":[251],"performing":[260],"presence":[265,503],"select":[269],"heparin-resistant":[271],"variants.":[272],"Characterisation":[273],"selected":[275],"revealed":[277,474],"single":[280],"round":[281,329],"had":[284],"variant":[288,355],"(P550S,":[289],"T588S)":[290],"4-fold":[293],"resistance.":[297],"IC50":[299,367],"increased":[301,368],"0.012U/ml":[303],"0.050U/ml":[306],"heparin.":[307,437],"study":[309,418],"followed":[313,333],"phytate":[316,371,377,435,465,516,572],"screen,":[317],"rounds":[321,349],"were":[324,399,448,525],"performed":[325,449],"initial":[328],"re-diversification":[335],"(recombination)":[336],"using":[341],"staggered":[343],"extension":[344],"process":[345],"(StEP).":[346],"yielded":[352],"2-fold":[358],"phytate-resistance":[361],"compared":[362],"wild-type,":[365],"360\u03bcM":[370],"700\u03bcM":[373],"best":[376],"(P685S,":[379],"M761V,":[380],"A814T)":[381],"further":[383,446],"characterised":[384],"it":[386],"catalytic":[391],"activity,":[392],"thermostability":[393],"fidelity":[395],"comparable":[400],"wildtype":[403],"enzyme.":[404],"position":[406],"resistance-conferring":[408],"mutations":[409],"evolved":[415],"provided":[419],"some":[420],"evidence":[421,480],"inhibitors\u2019":[424],"predicted":[425],"modes":[426],"case":[431],"2":[432],"both":[434],"As":[438],"phytate\u2019s":[439],"poorly":[444],"investigations":[447],"elucidate":[451],"role":[453],"inhibition.":[456],"thorough":[458],"investigation":[459],"into":[460,562],"importance":[462],"relative":[464],"levels":[468],"conducted":[472],"first":[477],"time":[478],"convincing":[479],"primary":[483],"phytatemediated":[486],"chelation.":[491],"Further":[492],"work":[493],"led":[494],"successful":[497],"crystallisation":[498],"although":[506],"subsequent":[507],"X-ray":[508],"diffraction":[509],"data":[510],"2.5\u00c5":[512],"did":[513],"reveal":[515],"bound":[517],"within":[518],"structure.":[521],"Site-directed":[522],"studies":[524],"probe":[528],"cross-over":[529],"between":[530],"phytate-conferring":[533],"mutations.":[534],"Thus,":[535],"addition":[537],"providing":[539],"valuable":[540],"information":[541],"potential":[548],"application":[549],"fecal-based":[551],"tests,":[554],"project":[556],"has":[557],"begun":[558],"provide":[560],"insight":[561],"fundamental":[564],"aspects":[565],"inhibitor.":[578]},"counts_by_year":[],"updated_date":"2026-09-29T14:09:16.559479","created_date":"2016-06-24T00:00:00"}
