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Article: Organization of the shrimp vitellogenin gene: Evidence of multiple genes and tissue specific expression by the ovary and hepatopancreas

TitleOrganization of the shrimp vitellogenin gene: Evidence of multiple genes and tissue specific expression by the ovary and hepatopancreas
Authors
KeywordsGene organization
Shrimp
Vitellogenin
Issue Date2003
PublisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/gene
Citation
Gene, 2003, v. 303 n. 1-2, p. 99-109 How to Cite?
AbstractVitellogenin is the major egg yolk protein synthesized in female shrimp during gonad maturation. Although there are several reports for the cloning of vitellogenin complementary DNA (cDNA) in different crustaceans, little is known of the gene organization of this protein. This study reports the first cloning and characterization of a full-length gene encoding the vitellogenin precursor from the shrimp Metapenaeus ensis. By genomic DNA library screening, six different lambda clones were isolated using shrimp partial gene sequence as probe. Initial DNA sequence determination revealed that these clones are derived from different genes with coding sequence similar to other crustacean vitellogenins. Two of these clones were used for further analysis. One of the lambda clones (λ3.3) carries most of the coding sequence that correspond to the M. ensis vitellogenin gene (MeVg1) and the other clone (λ8.3) carries a smaller portion of the coding sequence of a different vitellogenin gene (MeVg2). The λ3.3 clone was chosen for further characterization. To clone the remaining 5′ end upstream promoter region, 5′ untranslated region and the remaining coding sequence of MeVg1, a polymerase chain reaction (PCR)-based gene walking approach was used. Subsequently, a PCR clone with overlapping sequence identical to the genomic clone was obtained and the organization of MeVg1 gene was constructed. The MeVg1 gene consists of 15 exons and 14 introns spanning approximately 10 kb. Several potential cleavage sites were identified from the deduced vitellogenin precursor. Cleaving of the precursor in these sites would result in the production of several vitellogenin subunits. To clone the cDNA for the vitellogenin, 5′ and 3′ rapid amplification of cDNA ends was performed using ovary cDNA of the shrimp. A 4.4 kb 5′ cDNA clone and a 4 kb 3′ end cDNA clone were isolated. The size of the reconstructed cDNA for M. ensis Vg is 7.97 kb and consists of the longest open reading frame of 7776 bp. Unlike the vitellogenin precursor of most insects and vertebrates, the deduced vitellogenin precursor lacks the polyserine domain important for receptor-mediated endocytosis. Phylogenetic analysis revealed a closer relationship of the MeVg1 with other crustacean vitellogenins but distantly related to other invertebrate and vertebrate vitellogenins. By reverse transcription-PCR, we have demonstrated that the shrimp MeVg1 gene is expressed only in the ovary and hepatopancreas while the MeVg2 gene is expressed exclusively in the hepatopancreas. In conclusion, the shrimp ovary also contribute significantly in the production of vitellogenin at transcription level and the gene organization of the shrimp protein may provide an insight in the evolution of this group of important proteins. © 2002 Elsevier Science B.V. All rights reserved.
Persistent Identifierhttp://hdl.handle.net/10722/84851
ISSN
2015 Impact Factor: 2.319
2015 SCImago Journal Rankings: 1.043
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorTsang, WSen_HK
dc.contributor.authorQuackenbush, LSen_HK
dc.contributor.authorChow, BKCen_HK
dc.contributor.authorTiu, SHKen_HK
dc.contributor.authorHe, JGen_HK
dc.contributor.authorChan, SMen_HK
dc.date.accessioned2010-09-06T08:57:51Z-
dc.date.available2010-09-06T08:57:51Z-
dc.date.issued2003en_HK
dc.identifier.citationGene, 2003, v. 303 n. 1-2, p. 99-109en_HK
dc.identifier.issn0378-1119en_HK
dc.identifier.urihttp://hdl.handle.net/10722/84851-
dc.description.abstractVitellogenin is the major egg yolk protein synthesized in female shrimp during gonad maturation. Although there are several reports for the cloning of vitellogenin complementary DNA (cDNA) in different crustaceans, little is known of the gene organization of this protein. This study reports the first cloning and characterization of a full-length gene encoding the vitellogenin precursor from the shrimp Metapenaeus ensis. By genomic DNA library screening, six different lambda clones were isolated using shrimp partial gene sequence as probe. Initial DNA sequence determination revealed that these clones are derived from different genes with coding sequence similar to other crustacean vitellogenins. Two of these clones were used for further analysis. One of the lambda clones (λ3.3) carries most of the coding sequence that correspond to the M. ensis vitellogenin gene (MeVg1) and the other clone (λ8.3) carries a smaller portion of the coding sequence of a different vitellogenin gene (MeVg2). The λ3.3 clone was chosen for further characterization. To clone the remaining 5′ end upstream promoter region, 5′ untranslated region and the remaining coding sequence of MeVg1, a polymerase chain reaction (PCR)-based gene walking approach was used. Subsequently, a PCR clone with overlapping sequence identical to the genomic clone was obtained and the organization of MeVg1 gene was constructed. The MeVg1 gene consists of 15 exons and 14 introns spanning approximately 10 kb. Several potential cleavage sites were identified from the deduced vitellogenin precursor. Cleaving of the precursor in these sites would result in the production of several vitellogenin subunits. To clone the cDNA for the vitellogenin, 5′ and 3′ rapid amplification of cDNA ends was performed using ovary cDNA of the shrimp. A 4.4 kb 5′ cDNA clone and a 4 kb 3′ end cDNA clone were isolated. The size of the reconstructed cDNA for M. ensis Vg is 7.97 kb and consists of the longest open reading frame of 7776 bp. Unlike the vitellogenin precursor of most insects and vertebrates, the deduced vitellogenin precursor lacks the polyserine domain important for receptor-mediated endocytosis. Phylogenetic analysis revealed a closer relationship of the MeVg1 with other crustacean vitellogenins but distantly related to other invertebrate and vertebrate vitellogenins. By reverse transcription-PCR, we have demonstrated that the shrimp MeVg1 gene is expressed only in the ovary and hepatopancreas while the MeVg2 gene is expressed exclusively in the hepatopancreas. In conclusion, the shrimp ovary also contribute significantly in the production of vitellogenin at transcription level and the gene organization of the shrimp protein may provide an insight in the evolution of this group of important proteins. © 2002 Elsevier Science B.V. All rights reserved.en_HK
dc.languageengen_HK
dc.publisherElsevier BV. The Journal's web site is located at http://www.elsevier.com/locate/geneen_HK
dc.relation.ispartofGeneen_HK
dc.rightsGene. Copyright © Elsevier BV.en_HK
dc.subjectGene organizationen_HK
dc.subjectShrimpen_HK
dc.subjectVitellogeninen_HK
dc.titleOrganization of the shrimp vitellogenin gene: Evidence of multiple genes and tissue specific expression by the ovary and hepatopancreasen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=0378-1119&volume=303&spage=99&epage=109&date=2003&atitle=Organization+of+the+shrimp+vitellogenin+gene:+evidence+of+multiple+genes+and+tissue+specific+expression+by+the+ovary+and+hepatopancreasen_HK
dc.identifier.emailChow, BKC: bkcc@hku.hken_HK
dc.identifier.authorityChow, BKC=rp00681en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/S0378-1119(02)01139-3en_HK
dc.identifier.pmid12559571en_HK
dc.identifier.scopuseid_2-s2.0-0037448614en_HK
dc.identifier.hkuros75830en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-0037448614&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume303en_HK
dc.identifier.issue1-2en_HK
dc.identifier.spage99en_HK
dc.identifier.epage109en_HK
dc.identifier.isiWOS:000181124200010-
dc.publisher.placeNetherlandsen_HK
dc.identifier.scopusauthoridTsang, WS=7201558532en_HK
dc.identifier.scopusauthoridQuackenbush, LS=7004083800en_HK
dc.identifier.scopusauthoridChow, BKC=7102826193en_HK
dc.identifier.scopusauthoridTiu, SHK=8509766700en_HK
dc.identifier.scopusauthoridHe, JG=35487616500en_HK
dc.identifier.scopusauthoridChan, SM=7404255669en_HK

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