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Article: Molecular cloning, expression of orange-spotted grouper goose-type lysozyme cDNA, and lytic activity of its recombinant protein

TitleMolecular cloning, expression of orange-spotted grouper goose-type lysozyme cDNA, and lytic activity of its recombinant protein
Authors
KeywordsGoose-type lysozyme
Orange-spotted grouper
Lytic activity
cDNA library
Issue Date2003
PublisherInter-Research. The Journal's web site is located at http://www.int-res.com/journals/dao/index.html
Citation
Diseases of Aquatic Organisms, 2003, v. 55 n. 2, p. 117-123 How to Cite?
AbstractLysozyme acts as a non-specific innate-immunity molecule against the invasion of bacteria pathogens. A leukocyte cDNA library of orange-spotted grouper Epinephelus coioides was constructed and the goose-type (g-type) lysozyme cDNA was isolated. The complete cDNA consists of an open reading frame of 585 bp encoding a protein of 194 amino acids. This protein shows a 72.2% amino acid sequence identity with the flounder g-type lysozyme. Similar to most other species, the glu catalytic residue in g-type lysozymes of the grouper is conserved. Furthermore, like the flounder and carp, the 4 conserved cysteine residues identified in avian and mammalian g-type lysozymes were also absent from the grouper. Northern blot analysis indicated that the g-type lysozyme was expressed in intestine, liver, spleen, anterior kidney, posterior kidney, heart, gill, muscle and leukocytes. In addition, RT-PCR analysis detected the g-type lysozyme transcripts in the stomach, brain and ovary. When an orange-spotted grouper was injected with Vibrio alginolyticus, the number of lysozyme mRNA transcripts detected in the stomach, spleen, anterior kidney, posterior kidney, heart, brain and leucocytes increased 72 h after injection. Recombinant grouper g-type lysozyme produced in the Escherichia coli expression system showed lytic activity against Micrococcus lysodeikticus, V. alginolyticus from Epinephelus fario, V. vulnificus from culture water, Aeromonas hydrophila from soft-shell turtle, A. hydrophila from goldfish and V. parahaemolyticus, Pseudomonas fluorescens and V. fluvialis from culture water.
Persistent Identifierhttp://hdl.handle.net/10722/57392
ISSN
2023 Impact Factor: 1.1
2023 SCImago Journal Rankings: 0.438
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorYin, ZXen_HK
dc.contributor.authorHe, JGen_HK
dc.contributor.authorDeng, WXen_HK
dc.contributor.authorChan, SMen_HK
dc.date.accessioned2010-04-12T01:35:02Z-
dc.date.available2010-04-12T01:35:02Z-
dc.date.issued2003en_HK
dc.identifier.citationDiseases of Aquatic Organisms, 2003, v. 55 n. 2, p. 117-123en_HK
dc.identifier.issn0177-5103en_HK
dc.identifier.urihttp://hdl.handle.net/10722/57392-
dc.description.abstractLysozyme acts as a non-specific innate-immunity molecule against the invasion of bacteria pathogens. A leukocyte cDNA library of orange-spotted grouper Epinephelus coioides was constructed and the goose-type (g-type) lysozyme cDNA was isolated. The complete cDNA consists of an open reading frame of 585 bp encoding a protein of 194 amino acids. This protein shows a 72.2% amino acid sequence identity with the flounder g-type lysozyme. Similar to most other species, the glu catalytic residue in g-type lysozymes of the grouper is conserved. Furthermore, like the flounder and carp, the 4 conserved cysteine residues identified in avian and mammalian g-type lysozymes were also absent from the grouper. Northern blot analysis indicated that the g-type lysozyme was expressed in intestine, liver, spleen, anterior kidney, posterior kidney, heart, gill, muscle and leukocytes. In addition, RT-PCR analysis detected the g-type lysozyme transcripts in the stomach, brain and ovary. When an orange-spotted grouper was injected with Vibrio alginolyticus, the number of lysozyme mRNA transcripts detected in the stomach, spleen, anterior kidney, posterior kidney, heart, brain and leucocytes increased 72 h after injection. Recombinant grouper g-type lysozyme produced in the Escherichia coli expression system showed lytic activity against Micrococcus lysodeikticus, V. alginolyticus from Epinephelus fario, V. vulnificus from culture water, Aeromonas hydrophila from soft-shell turtle, A. hydrophila from goldfish and V. parahaemolyticus, Pseudomonas fluorescens and V. fluvialis from culture water.en_HK
dc.languageengen_HK
dc.publisherInter-Research. The Journal's web site is located at http://www.int-res.com/journals/dao/index.htmlen_HK
dc.rightsDiseases of Aquatic Organisms. Copyright © Inter-Research.en_HK
dc.subjectGoose-type lysozymeen_HK
dc.subjectOrange-spotted grouperen_HK
dc.subjectLytic activityen_HK
dc.subjectcDNA libraryen_HK
dc.titleMolecular cloning, expression of orange-spotted grouper goose-type lysozyme cDNA, and lytic activity of its recombinant proteinen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0177-5103&volume=55&issue=2&spage=117&epage=123&date=2003&atitle=Molecular+cloning,+expression+of+orange-spotted+grouper+goose-type+lysozyme+cDNA,+and+lytic+activity+of+its+recombinant+proteinen_HK
dc.identifier.emailChan, SM: chansm@hkucc.hku.hken_HK
dc.description.naturepublished_or_final_versionen_HK
dc.identifier.doi10.3354/dao055117en_HK
dc.identifier.pmid12911059en_HK
dc.identifier.scopuseid_2-s2.0-0042267811-
dc.identifier.hkuros85843-
dc.identifier.isiWOS:000185151200005-
dc.identifier.issnl0177-5103-

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