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Organization of the Shrimp (Metapenaeus Ensis) Vitellogenin Gene: Evidence for Multiple Genes and Ovary Expression

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This dissertation, "Organization of the Shrimp (Metapenaeus Ensis) Vitellogenin Gene: Evidence for Multiple Genes and Ovary Expression" by Wing-sze, Tsang, 曾詠思, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any wa This dissertation, "Organization of the Shrimp (Metapenaeus Ensis) Vitellogenin Gene: Evidence for Multiple Genes and Ovary Expression" by Wing-sze, Tsang, 曾詠思, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled ORGANIZATION OF THE SHRIMP (Metapenaeus ensis) VITELLOGENIN GENE: EVIDENCE FOR MULTIPLE GENES AND OVARY EXPRESSION Submitted by Tsang Wing Sze for the degree of Master of Philosophy at The University of Hong Kong in December 2002 Vitellogenin (Vg) is the major egg yolk protein that plays an important role in the reproduction of oviparous animals. In crustaceans, there are many conflicting reports for the synthesis site of Vg. The possible synthetic sites include the subepidermal adipose tissue, the hepatopancreas and the ovary. The objectives of this study were to characterize the gene encoding the major egg yolk protein Vg in shrimp and to investigate the possible sites for Vg synthesis by molecular approaches. After library screening, several Lambda genomic DNA clones carrying the partial Vg gene were isolated and purified. By partial sequencing of different Lambda genomic DNA clones and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) cloning, this study has provided evidence that Vg is encoded by multiple genes in the shrimp Metapenaeus ensis. In addition, the full-length genomic and cDNA sequence of one of the Vg genes (MeVg1) was cloned and characterized. The MeVg1 gene is 10. 4 kb in size and consists of 15 exons interrupted by 14 introns. The open reading frame (ORF) of the Vg cDNA (MeVg1) is 7.779 kb long encoding 2,592 amino acid residues. It consists of 47 bp of 5' UTR and 203 bp of 3' UTR. The amino acid sequence of MeVg1 shows high amino-acid similarity to other crustacean species. Phylogenetic analysis of different Vgs indicate that the malacostracean crustaceans form a sister group among themselves and that the group diverges from the clade of arthropods. By Northern blot and RT-PCR analyses, two Vg genes (MeVg1 and MeVg2) were identified. The expression patterns of these genes in the hepatopancreas and ovary are different. During the reproductive phase of the female, MeVg1 mRNA transcripts can be detected in both the hepatopancreas and ovary whereas MeVg2 mRNA transcripts can be detected in the hepatopancreas only. The expression level of MeVg1 is the highest at stage III in the hepatopancreas and the highest at stage V in the ovary. To summarize, this is the first report on the full-length sequence and gene organization of Vg in crustaceans. This study also provides evidence that the shrimp Vg is encoded by a small number of genes. Due to the complicated structure of the MeVg1 gene and the existence of multiple genes in the shrimp genome, the expression patterns of the vitellogenin genes in shrimp and the confirmation for the major synthesis sites need further investigation. The results of this study also indicate that the evolution and expression patterns of vitellogenin genes in shrimp are much more complicated than previously believed. DOI: 10.5353/th_b2663568 Subjects: Lipoproteins Gene expression Shrimps - Genetics


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This dissertation, "Organization of the Shrimp (Metapenaeus Ensis) Vitellogenin Gene: Evidence for Multiple Genes and Ovary Expression" by Wing-sze, Tsang, 曾詠思, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any wa This dissertation, "Organization of the Shrimp (Metapenaeus Ensis) Vitellogenin Gene: Evidence for Multiple Genes and Ovary Expression" by Wing-sze, Tsang, 曾詠思, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled ORGANIZATION OF THE SHRIMP (Metapenaeus ensis) VITELLOGENIN GENE: EVIDENCE FOR MULTIPLE GENES AND OVARY EXPRESSION Submitted by Tsang Wing Sze for the degree of Master of Philosophy at The University of Hong Kong in December 2002 Vitellogenin (Vg) is the major egg yolk protein that plays an important role in the reproduction of oviparous animals. In crustaceans, there are many conflicting reports for the synthesis site of Vg. The possible synthetic sites include the subepidermal adipose tissue, the hepatopancreas and the ovary. The objectives of this study were to characterize the gene encoding the major egg yolk protein Vg in shrimp and to investigate the possible sites for Vg synthesis by molecular approaches. After library screening, several Lambda genomic DNA clones carrying the partial Vg gene were isolated and purified. By partial sequencing of different Lambda genomic DNA clones and Reverse Transcription-Polymerase Chain Reaction (RT-PCR) cloning, this study has provided evidence that Vg is encoded by multiple genes in the shrimp Metapenaeus ensis. In addition, the full-length genomic and cDNA sequence of one of the Vg genes (MeVg1) was cloned and characterized. The MeVg1 gene is 10. 4 kb in size and consists of 15 exons interrupted by 14 introns. The open reading frame (ORF) of the Vg cDNA (MeVg1) is 7.779 kb long encoding 2,592 amino acid residues. It consists of 47 bp of 5' UTR and 203 bp of 3' UTR. The amino acid sequence of MeVg1 shows high amino-acid similarity to other crustacean species. Phylogenetic analysis of different Vgs indicate that the malacostracean crustaceans form a sister group among themselves and that the group diverges from the clade of arthropods. By Northern blot and RT-PCR analyses, two Vg genes (MeVg1 and MeVg2) were identified. The expression patterns of these genes in the hepatopancreas and ovary are different. During the reproductive phase of the female, MeVg1 mRNA transcripts can be detected in both the hepatopancreas and ovary whereas MeVg2 mRNA transcripts can be detected in the hepatopancreas only. The expression level of MeVg1 is the highest at stage III in the hepatopancreas and the highest at stage V in the ovary. To summarize, this is the first report on the full-length sequence and gene organization of Vg in crustaceans. This study also provides evidence that the shrimp Vg is encoded by a small number of genes. Due to the complicated structure of the MeVg1 gene and the existence of multiple genes in the shrimp genome, the expression patterns of the vitellogenin genes in shrimp and the confirmation for the major synthesis sites need further investigation. The results of this study also indicate that the evolution and expression patterns of vitellogenin genes in shrimp are much more complicated than previously believed. DOI: 10.5353/th_b2663568 Subjects: Lipoproteins Gene expression Shrimps - Genetics

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