Genetic, biochemical characterization and mutagenesis of the chromosomal class A β-lactamase of Raoultella (formerly Klebsiella) terrigena
Chromosomal class A β-lactamases have been characterized in Raoultella ornithinolytica and Raoultella planticola. The purpose of this study was to characterize that of Raoultella terrigena. The blaTER-1 gene of R. terrigena strain ATCC33257T was cloned (pACter-1) and sequenced. It was then used to d...
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| Veröffentlicht in: | Pathologie biologie (Paris) 2015-09, Vol.63 (4-5), p.158-163 |
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| creator | Walckenaer, E. Delmas, J. Leflon-Guibout, V. Bonnet, R. Nicolas-Chanoine, M.-H. |
| description | Chromosomal class A β-lactamases have been characterized in Raoultella ornithinolytica and Raoultella planticola. The purpose of this study was to characterize that of Raoultella terrigena.
The blaTER-1 gene of R. terrigena strain ATCC33257T was cloned (pACter-1) and sequenced. It was then used to detect the bla gene of strains BM 85 01 095 and SB2796. The hypermutable Escherichia coli strain AB1157 mutS::Tn10 was transformed with pACter-1 and mutants growing on plates containing>2mg/L ceftazidime were studied. Notably, the impact of mutations only observed in the promoter region on β-lactam resistance was assessed by site-directed mutagenesis experiments.
R. terrigena strains ATCC33257T and BM 85 01 095 had the same bla gene and deduced protein (TER-1) whereas there were 3 substitutions in those of strain SB2796 (TER-2). Class A β-lactamases TER showed 78%, 69.9% and 38.7% identity with PLA or ORN, TEM-1 and KOXY, respectively. Compared with TEM-1, TER-1 and TER-2 showed 2 particular substitutions, Leu75Pro and Glu240Asn demonstrated to be involved in the inherent β-lactam resistance profile of R. terrigena. TER-1 (pI of 7.6) had a high activity against penicillin G and a significantly low one against amoxicillin. Substitution G/T observed in the -35 region of the blaTER gene harbored by strains growing in the presence of≥2mg/L ceftazidime was shown to be responsible for this growth.
TER is a new class A β-lactamase belonging to functional group 2b.
Les β-lactamases chromosomiques de classe A de Raoultella ornithinolytica et Raoultella planticola ont été caractérisées. Le but de la présente étude était de caractériser celle de Raoultella terrigena.
Le gène blaTER-1 de la souche de R. terrigena ATCC33257T a été cloné (pACter-1), séquencé, puis utilisé pour la détection du gène bla des souches de R. terrigena BM 85 01 095 et SB2796. La souche hypermutatrice de Escherichia coli AB1157 (mutS::Tn10) a été transformée avec le plasmide pACter-1 et les mutants cultivant en présence de>2mg/L de ceftazidime ont été étudiés.
Les séquences du gène et de la protéine déduite (TER-1) sont identiques chez les souches ATCC33257T et BM 85 01 095 tandis que 3 substitutions sont observées dans la protéine déduite (TER-2) chez la souche SB2796. La β-lactamase TER de classe A présente 78 %, 69,9 % et 38,7 % d’identité avec les β-lactamases PLA ou ORN, TEM-1 et KOXY, respectivement. Comparées à TEM-1, TER-1 et TER-2 ont 2 substitutions particulières, Leu75Pro et Glu240Asn, dont |
| doi_str_mv | 10.1016/j.patbio.2015.05.002 |
| format | Article |
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The blaTER-1 gene of R. terrigena strain ATCC33257T was cloned (pACter-1) and sequenced. It was then used to detect the bla gene of strains BM 85 01 095 and SB2796. The hypermutable Escherichia coli strain AB1157 mutS::Tn10 was transformed with pACter-1 and mutants growing on plates containing>2mg/L ceftazidime were studied. Notably, the impact of mutations only observed in the promoter region on β-lactam resistance was assessed by site-directed mutagenesis experiments.
R. terrigena strains ATCC33257T and BM 85 01 095 had the same bla gene and deduced protein (TER-1) whereas there were 3 substitutions in those of strain SB2796 (TER-2). Class A β-lactamases TER showed 78%, 69.9% and 38.7% identity with PLA or ORN, TEM-1 and KOXY, respectively. Compared with TEM-1, TER-1 and TER-2 showed 2 particular substitutions, Leu75Pro and Glu240Asn demonstrated to be involved in the inherent β-lactam resistance profile of R. terrigena. TER-1 (pI of 7.6) had a high activity against penicillin G and a significantly low one against amoxicillin. Substitution G/T observed in the -35 region of the blaTER gene harbored by strains growing in the presence of≥2mg/L ceftazidime was shown to be responsible for this growth.
TER is a new class A β-lactamase belonging to functional group 2b.
Les β-lactamases chromosomiques de classe A de Raoultella ornithinolytica et Raoultella planticola ont été caractérisées. Le but de la présente étude était de caractériser celle de Raoultella terrigena.
Le gène blaTER-1 de la souche de R. terrigena ATCC33257T a été cloné (pACter-1), séquencé, puis utilisé pour la détection du gène bla des souches de R. terrigena BM 85 01 095 et SB2796. La souche hypermutatrice de Escherichia coli AB1157 (mutS::Tn10) a été transformée avec le plasmide pACter-1 et les mutants cultivant en présence de>2mg/L de ceftazidime ont été étudiés.
Les séquences du gène et de la protéine déduite (TER-1) sont identiques chez les souches ATCC33257T et BM 85 01 095 tandis que 3 substitutions sont observées dans la protéine déduite (TER-2) chez la souche SB2796. La β-lactamase TER de classe A présente 78 %, 69,9 % et 38,7 % d’identité avec les β-lactamases PLA ou ORN, TEM-1 et KOXY, respectivement. Comparées à TEM-1, TER-1 et TER-2 ont 2 substitutions particulières, Leu75Pro et Glu240Asn, dont l’implication dans le profil de résistance de R. terrigena a été démontrée dans la présente étude. TER-1 (pI of 7.6) hydrolyse fortement la pénicilline G et significativement moins l’amoxicilline. La substitution G/T observée dans la région -35 du gène blaTER chez les mutants cultivant en présence de≥2mg/L de ceftazidime est responsable de cette croissance.
TER est une nouvelle β-lactamase de classe A appartenant au groupe fonctionnel 2b.</description><identifier>ISSN: 0369-8114</identifier><identifier>EISSN: 1768-3114</identifier><identifier>DOI: 10.1016/j.patbio.2015.05.002</identifier><identifier>PMID: 26092758</identifier><language>eng</language><publisher>France: Elsevier SAS</publisher><subject>Amino Acid Sequence ; Amino Acid Substitution ; Anti-Bacterial Agents - pharmacology ; Bacterial Proteins - genetics ; Bacterial Proteins - metabolism ; beta-Lactam Resistance - genetics ; beta-Lactamases - classification ; beta-Lactamases - genetics ; beta-Lactamases - metabolism ; beta-Lactams - pharmacology ; bla gene ; Ceftazidime - pharmacology ; Chromosomes, Bacterial ; Cloning, Molecular ; DNA Transposable Elements ; Drug Resistance, Multiple, Bacterial - genetics ; Enterobacteriaceae - classification ; Enterobacteriaceae - enzymology ; Enterobacteriaceae - genetics ; Escherichia coli ; Genes, Bacterial ; Gène bla ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Promoter Regions, Genetic ; Promoteur fort ; Raoultella terrigena ; Sequence Alignment ; Sequence Homology, Amino Acid ; Strength promoter</subject><ispartof>Pathologie biologie (Paris), 2015-09, Vol.63 (4-5), p.158-163</ispartof><rights>2015</rights><rights>Copyright © 2015. Published by Elsevier SAS.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c432t-82605c7ea279672cd732224156f8ead9b38e3ab74650e8a610c74fbe0b1ff1cb3</citedby><cites>FETCH-LOGICAL-c432t-82605c7ea279672cd732224156f8ead9b38e3ab74650e8a610c74fbe0b1ff1cb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0369811415000498$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26092758$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Walckenaer, E.</creatorcontrib><creatorcontrib>Delmas, J.</creatorcontrib><creatorcontrib>Leflon-Guibout, V.</creatorcontrib><creatorcontrib>Bonnet, R.</creatorcontrib><creatorcontrib>Nicolas-Chanoine, M.-H.</creatorcontrib><title>Genetic, biochemical characterization and mutagenesis of the chromosomal class A β-lactamase of Raoultella (formerly Klebsiella) terrigena</title><title>Pathologie biologie (Paris)</title><addtitle>Pathol Biol (Paris)</addtitle><description>Chromosomal class A β-lactamases have been characterized in Raoultella ornithinolytica and Raoultella planticola. The purpose of this study was to characterize that of Raoultella terrigena.
The blaTER-1 gene of R. terrigena strain ATCC33257T was cloned (pACter-1) and sequenced. It was then used to detect the bla gene of strains BM 85 01 095 and SB2796. The hypermutable Escherichia coli strain AB1157 mutS::Tn10 was transformed with pACter-1 and mutants growing on plates containing>2mg/L ceftazidime were studied. Notably, the impact of mutations only observed in the promoter region on β-lactam resistance was assessed by site-directed mutagenesis experiments.
R. terrigena strains ATCC33257T and BM 85 01 095 had the same bla gene and deduced protein (TER-1) whereas there were 3 substitutions in those of strain SB2796 (TER-2). Class A β-lactamases TER showed 78%, 69.9% and 38.7% identity with PLA or ORN, TEM-1 and KOXY, respectively. Compared with TEM-1, TER-1 and TER-2 showed 2 particular substitutions, Leu75Pro and Glu240Asn demonstrated to be involved in the inherent β-lactam resistance profile of R. terrigena. TER-1 (pI of 7.6) had a high activity against penicillin G and a significantly low one against amoxicillin. Substitution G/T observed in the -35 region of the blaTER gene harbored by strains growing in the presence of≥2mg/L ceftazidime was shown to be responsible for this growth.
TER is a new class A β-lactamase belonging to functional group 2b.
Les β-lactamases chromosomiques de classe A de Raoultella ornithinolytica et Raoultella planticola ont été caractérisées. Le but de la présente étude était de caractériser celle de Raoultella terrigena.
Le gène blaTER-1 de la souche de R. terrigena ATCC33257T a été cloné (pACter-1), séquencé, puis utilisé pour la détection du gène bla des souches de R. terrigena BM 85 01 095 et SB2796. La souche hypermutatrice de Escherichia coli AB1157 (mutS::Tn10) a été transformée avec le plasmide pACter-1 et les mutants cultivant en présence de>2mg/L de ceftazidime ont été étudiés.
Les séquences du gène et de la protéine déduite (TER-1) sont identiques chez les souches ATCC33257T et BM 85 01 095 tandis que 3 substitutions sont observées dans la protéine déduite (TER-2) chez la souche SB2796. La β-lactamase TER de classe A présente 78 %, 69,9 % et 38,7 % d’identité avec les β-lactamases PLA ou ORN, TEM-1 et KOXY, respectivement. Comparées à TEM-1, TER-1 et TER-2 ont 2 substitutions particulières, Leu75Pro et Glu240Asn, dont l’implication dans le profil de résistance de R. terrigena a été démontrée dans la présente étude. TER-1 (pI of 7.6) hydrolyse fortement la pénicilline G et significativement moins l’amoxicilline. La substitution G/T observée dans la région -35 du gène blaTER chez les mutants cultivant en présence de≥2mg/L de ceftazidime est responsable de cette croissance.
TER est une nouvelle β-lactamase de classe A appartenant au groupe fonctionnel 2b.</description><subject>Amino Acid Sequence</subject><subject>Amino Acid Substitution</subject><subject>Anti-Bacterial Agents - pharmacology</subject><subject>Bacterial Proteins - genetics</subject><subject>Bacterial Proteins - metabolism</subject><subject>beta-Lactam Resistance - genetics</subject><subject>beta-Lactamases - classification</subject><subject>beta-Lactamases - genetics</subject><subject>beta-Lactamases - metabolism</subject><subject>beta-Lactams - pharmacology</subject><subject>bla gene</subject><subject>Ceftazidime - pharmacology</subject><subject>Chromosomes, Bacterial</subject><subject>Cloning, Molecular</subject><subject>DNA Transposable Elements</subject><subject>Drug Resistance, Multiple, Bacterial - genetics</subject><subject>Enterobacteriaceae - classification</subject><subject>Enterobacteriaceae - enzymology</subject><subject>Enterobacteriaceae - genetics</subject><subject>Escherichia coli</subject><subject>Genes, Bacterial</subject><subject>Gène bla</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Site-Directed</subject><subject>Promoter Regions, Genetic</subject><subject>Promoteur fort</subject><subject>Raoultella terrigena</subject><subject>Sequence Alignment</subject><subject>Sequence Homology, Amino Acid</subject><subject>Strength promoter</subject><issn>0369-8114</issn><issn>1768-3114</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kdFqFDEUhoNY7Fp9A5FcVuisSWYmmb0RSrFVWhBEr8OZzBk3SzJZk0yhfQXfpg_SZzLD1l4KBxLC95__5PyEvONszRmXH3frPeTehrVgvF2zUky8ICuuZFfVnDcvyYrVclN15X5MXqe0Y4wr3vBX5FhIthGq7VbkzxVOmK05o6WV2aK3Bhw1W4hgMkZ7D9mGicI0UD9n-FXoZBMNI81bLFwMPqTgF42DlOg5fXyoXNGCh4QL9x3C7DI6B_R0DNFjdHf02mGf7PL4gRabaEtjeEOORnAJ3z6dJ-Tn5ecfF1-qm29XXy_ObyrT1CJXXZm-NQpBqI1UwgyqFkI0vJVjhzBs-rrDGnrVyJZhB5Izo5qxR9bzceSmr0_I6aHvPobfM6asvU1mGWbCMCddttRKyZTiBW0OqIkhpYij3kfrId5pzvQSg97pQwx6iUGzUkwU2fsnh7n3ODyL_u29AJ8OAJZ_3lqMOhmLk8HBRjRZD8H-3-EvHP6d7g</recordid><startdate>20150901</startdate><enddate>20150901</enddate><creator>Walckenaer, E.</creator><creator>Delmas, J.</creator><creator>Leflon-Guibout, V.</creator><creator>Bonnet, R.</creator><creator>Nicolas-Chanoine, M.-H.</creator><general>Elsevier SAS</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20150901</creationdate><title>Genetic, biochemical characterization and mutagenesis of the chromosomal class A β-lactamase of Raoultella (formerly Klebsiella) terrigena</title><author>Walckenaer, E. ; Delmas, J. ; Leflon-Guibout, V. ; Bonnet, R. ; Nicolas-Chanoine, M.-H.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c432t-82605c7ea279672cd732224156f8ead9b38e3ab74650e8a610c74fbe0b1ff1cb3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Amino Acid Sequence</topic><topic>Amino Acid Substitution</topic><topic>Anti-Bacterial Agents - pharmacology</topic><topic>Bacterial Proteins - genetics</topic><topic>Bacterial Proteins - metabolism</topic><topic>beta-Lactam Resistance - genetics</topic><topic>beta-Lactamases - classification</topic><topic>beta-Lactamases - genetics</topic><topic>beta-Lactamases - metabolism</topic><topic>beta-Lactams - pharmacology</topic><topic>bla gene</topic><topic>Ceftazidime - pharmacology</topic><topic>Chromosomes, Bacterial</topic><topic>Cloning, Molecular</topic><topic>DNA Transposable Elements</topic><topic>Drug Resistance, Multiple, Bacterial - genetics</topic><topic>Enterobacteriaceae - classification</topic><topic>Enterobacteriaceae - enzymology</topic><topic>Enterobacteriaceae - genetics</topic><topic>Escherichia coli</topic><topic>Genes, Bacterial</topic><topic>Gène bla</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis, Site-Directed</topic><topic>Promoter Regions, Genetic</topic><topic>Promoteur fort</topic><topic>Raoultella terrigena</topic><topic>Sequence Alignment</topic><topic>Sequence Homology, Amino Acid</topic><topic>Strength promoter</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Walckenaer, E.</creatorcontrib><creatorcontrib>Delmas, J.</creatorcontrib><creatorcontrib>Leflon-Guibout, V.</creatorcontrib><creatorcontrib>Bonnet, R.</creatorcontrib><creatorcontrib>Nicolas-Chanoine, M.-H.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Pathologie biologie (Paris)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Walckenaer, E.</au><au>Delmas, J.</au><au>Leflon-Guibout, V.</au><au>Bonnet, R.</au><au>Nicolas-Chanoine, M.-H.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Genetic, biochemical characterization and mutagenesis of the chromosomal class A β-lactamase of Raoultella (formerly Klebsiella) terrigena</atitle><jtitle>Pathologie biologie (Paris)</jtitle><addtitle>Pathol Biol (Paris)</addtitle><date>2015-09-01</date><risdate>2015</risdate><volume>63</volume><issue>4-5</issue><spage>158</spage><epage>163</epage><pages>158-163</pages><issn>0369-8114</issn><eissn>1768-3114</eissn><abstract>Chromosomal class A β-lactamases have been characterized in Raoultella ornithinolytica and Raoultella planticola. The purpose of this study was to characterize that of Raoultella terrigena.
The blaTER-1 gene of R. terrigena strain ATCC33257T was cloned (pACter-1) and sequenced. It was then used to detect the bla gene of strains BM 85 01 095 and SB2796. The hypermutable Escherichia coli strain AB1157 mutS::Tn10 was transformed with pACter-1 and mutants growing on plates containing>2mg/L ceftazidime were studied. Notably, the impact of mutations only observed in the promoter region on β-lactam resistance was assessed by site-directed mutagenesis experiments.
R. terrigena strains ATCC33257T and BM 85 01 095 had the same bla gene and deduced protein (TER-1) whereas there were 3 substitutions in those of strain SB2796 (TER-2). Class A β-lactamases TER showed 78%, 69.9% and 38.7% identity with PLA or ORN, TEM-1 and KOXY, respectively. Compared with TEM-1, TER-1 and TER-2 showed 2 particular substitutions, Leu75Pro and Glu240Asn demonstrated to be involved in the inherent β-lactam resistance profile of R. terrigena. TER-1 (pI of 7.6) had a high activity against penicillin G and a significantly low one against amoxicillin. Substitution G/T observed in the -35 region of the blaTER gene harbored by strains growing in the presence of≥2mg/L ceftazidime was shown to be responsible for this growth.
TER is a new class A β-lactamase belonging to functional group 2b.
Les β-lactamases chromosomiques de classe A de Raoultella ornithinolytica et Raoultella planticola ont été caractérisées. Le but de la présente étude était de caractériser celle de Raoultella terrigena.
Le gène blaTER-1 de la souche de R. terrigena ATCC33257T a été cloné (pACter-1), séquencé, puis utilisé pour la détection du gène bla des souches de R. terrigena BM 85 01 095 et SB2796. La souche hypermutatrice de Escherichia coli AB1157 (mutS::Tn10) a été transformée avec le plasmide pACter-1 et les mutants cultivant en présence de>2mg/L de ceftazidime ont été étudiés.
Les séquences du gène et de la protéine déduite (TER-1) sont identiques chez les souches ATCC33257T et BM 85 01 095 tandis que 3 substitutions sont observées dans la protéine déduite (TER-2) chez la souche SB2796. La β-lactamase TER de classe A présente 78 %, 69,9 % et 38,7 % d’identité avec les β-lactamases PLA ou ORN, TEM-1 et KOXY, respectivement. Comparées à TEM-1, TER-1 et TER-2 ont 2 substitutions particulières, Leu75Pro et Glu240Asn, dont l’implication dans le profil de résistance de R. terrigena a été démontrée dans la présente étude. TER-1 (pI of 7.6) hydrolyse fortement la pénicilline G et significativement moins l’amoxicilline. La substitution G/T observée dans la région -35 du gène blaTER chez les mutants cultivant en présence de≥2mg/L de ceftazidime est responsable de cette croissance.
TER est une nouvelle β-lactamase de classe A appartenant au groupe fonctionnel 2b.</abstract><cop>France</cop><pub>Elsevier SAS</pub><pmid>26092758</pmid><doi>10.1016/j.patbio.2015.05.002</doi><tpages>6</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0369-8114 |
| ispartof | Pathologie biologie (Paris), 2015-09, Vol.63 (4-5), p.158-163 |
| issn | 0369-8114 1768-3114 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_1715660771 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Amino Acid Sequence Amino Acid Substitution Anti-Bacterial Agents - pharmacology Bacterial Proteins - genetics Bacterial Proteins - metabolism beta-Lactam Resistance - genetics beta-Lactamases - classification beta-Lactamases - genetics beta-Lactamases - metabolism beta-Lactams - pharmacology bla gene Ceftazidime - pharmacology Chromosomes, Bacterial Cloning, Molecular DNA Transposable Elements Drug Resistance, Multiple, Bacterial - genetics Enterobacteriaceae - classification Enterobacteriaceae - enzymology Enterobacteriaceae - genetics Escherichia coli Genes, Bacterial Gène bla Molecular Sequence Data Mutagenesis, Site-Directed Promoter Regions, Genetic Promoteur fort Raoultella terrigena Sequence Alignment Sequence Homology, Amino Acid Strength promoter |
| title | Genetic, biochemical characterization and mutagenesis of the chromosomal class A β-lactamase of Raoultella (formerly Klebsiella) terrigena |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-04T18%3A05%3A59IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Genetic,%20biochemical%20characterization%20and%20mutagenesis%20of%20the%20chromosomal%20class%20A%20%CE%B2-lactamase%20of%20Raoultella%20(formerly%20Klebsiella)%20terrigena&rft.jtitle=Pathologie%20biologie%20(Paris)&rft.au=Walckenaer,%20E.&rft.date=2015-09-01&rft.volume=63&rft.issue=4-5&rft.spage=158&rft.epage=163&rft.pages=158-163&rft.issn=0369-8114&rft.eissn=1768-3114&rft_id=info:doi/10.1016/j.patbio.2015.05.002&rft_dat=%3Cproquest_cross%3E1715660771%3C/proquest_cross%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1715660771&rft_id=info:pmid/26092758&rft_els_id=S0369811415000498&rfr_iscdi=true |