Molecular marine biology and biotechnology最新文献

{"title":"Nucleotide sequence of the promoter region of Sparus aurata insulin-like growth factor I gene and expression of IGF-I in eggs and embryos.","authors":"B Fukenstein,&nbsp;R Shemer,&nbsp;R Amuly,&nbsp;I Cohen","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>A genomic fragment of 3.1 kb, containing the promoter region of Sparus aurata insulin-like growth factor I (IGF-I) gene has been cloned and sequenced. This fragment contains exon 1 and exon 2 of Sparus aurata IGF-I gene. These two exons are identical in organization to the reported chum salmon IGF-I gene. Exon 1 contains 5' untranslated region and part of the signal peptide. Exon 2 codes for part of the signal peptide and part of the B domain. Nested polymerase chain reaction (PCR) revealed that a fragment was amplified from liver RNA when a common primer in the translated region and a primer located between 375 and 395 nucleotides upstream of the first methionine were used. No such amplification was obtained when the primer was located between 414 and 434 nucleotides upstream of the first methionine, suggesting that the first exon in Sparus IGF-I gene starts 400-410 nucleotides upstream of the first methionine. Expression of IGF-I mRNA was studied in Sparus aurata using reverse transcription (RT)-PCR. An amplified fragment was found in unfertilized eggs and in embryos 4, 8, and 12 hours after fertilization, when oligonucleotides specific for Sparus aurata IGF-I cDNA were used. A similar fragment was found when adult liver or one-day larval RNA were used. This fragment hybridized in a Southern blot to salmon IGF-I cDNA. These results demonstrate that the structure of IGF-I gene has been conserved in teleosts and IGF-I transcripts are present in fish during embryogenesis, probably of maternal origin.</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"5 1","pages":"43-51"},"PeriodicalIF":0.0,"publicationDate":"1996-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19835020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular analysis of a RAPD marker (B20) reveals two microsatellites and differential mRNA expression in Penaeus vannamei.","authors":"D K Garcia,&nbsp;A K Dhar,&nbsp;A Alcivar-Warren","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>We previously reported a population-specific DNA fragment (B20) in Penaeus vannamei shrimp, fragment found using the randomly amplified polymorphic DNA (RAPD) procedure, that was present in Population 2 but not in Populations 1 and 4. The specific objectives of this study were to clone and sequence this genetic marker, determine if all or part of this cloned sequence could be found in any of the other populations in which this marker could not be amplified, and examine if this marker represents a functional gene by examining the steady-state levels of mRNA expression using Northern blot hybridization. Sequence information of the 1259-bp B20 clone revealed two microsatellites and two candidate open reading frames. Although the entire B20 sequence could only be amplified in Population 2 (from Ecuador), Population 3 (a hybrid of Populations 1 and 2), and a few individuals from wild Ecuadorian shrimp samples, portions of the B20 DNA could be amplified in individuals from Populations 1, 2, 3, candidate Population 4, and wild Ecuadorian samples. These microsatellites vary in size between populations and families. Northern blot hybridization analysis using radiolabeled B20 probe detected two mRNA transcripts of approximately 1.5 and 2.0 kb. Expression data throughout development indicated that these transcripts were present at low levels in nauplii from two of the three crosses examined using broodstocks of Population 1. Higher levels were observed in postlarvae (PL) 6, PL8, and PL10 in one of the three crosses. Individuals from all crosses showed higher levels of expression in the juvenile tail muscle. The mRNA transcript levels were undetected in zoea 3, PL2, and PL4 stages of development and broodstock tail muscle. The levels of expression of B20 mRNA transcripts varied significantly between Populations 1, 2, 3, 4, and wild Ecuadorian individuals as well as between families and within individuals representative of seven families from Population 1. In summary, the B20 clone revealed the presence of two microsatellites that vary in size between populations. These microsatellites will be useful for estimating genetic diversity within and between populations, identifying family-specific markers, and mapping loci responsible for economically important traits in penaeid shrimp. The mRNA levels detected by the B20 clone showed differential expression during development, and the pattern of expression was influenced by the genetic background of the parental crosses used.</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"5 1","pages":"71-83"},"PeriodicalIF":0.0,"publicationDate":"1996-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19835023","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A diagnostic molecular marker for zebra mussels (Dreissena polymorpha) and potentially co-occurring bivalves: mitochondrial COI.","authors":"B S Baldwin,&nbsp;M Black,&nbsp;O Sanjur,&nbsp;R Gustafson,&nbsp;R A Lutz,&nbsp;R C Vrijenhoek","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>We report diagnostic differences in the nucleotide sequences of a 710-bp fragment of the mitochondrial cytochrome c oxidase subunit I gene (COI) from the zebra mussel (Dreissena polymorpha) and potentially co-occurring bivalves: the quagga mussel (Dreissena bugensis); the Asiatic clam (Corbicula fluminea), the dark false mussel (Mytilopsis leucophaeata), and the wedge clam (Rangia cuneata). The COI sequence of the deep-water \"profunda\" phenotype of the quagga mussel was nearly identical to that of shallow-water quagga mussels. Restriction fragment length polymorphisms (RFLPs) in this portion of COI produced species-specific differences in fragment numbers and sizes that could be used as diagnostic markers to distinguish the free-living larvae produced by these bivalves.</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"5 1","pages":"9-14"},"PeriodicalIF":0.0,"publicationDate":"1996-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19835018","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DNA sequence variation of mitochondrial large-subunit rRNA provides support for a two-subclass organization of the Anthozoa (Cnidaria).","authors":"S C France,&nbsp;P E Rosel,&nbsp;J E Agenbroad,&nbsp;L S Mullineaux,&nbsp;T D Kocher","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>We have sequenced a portion of the mitochondrial 16S rRNA gene from 29 species of Anthozoa, representing six orders of the subclasses Ceriantipatharia, Hexacorallia, and Octocorallia, with the focus on deep-seamount corals (> 500-m depth). We have detected significant length variation in the gene, with homologous gene fragments ranging from 545 bp in a shallow-water scleractinian coral to 911 bp in a deep-sea antipatharian black coral. The aligned sequences were divided into five regions: three high-identity sequence blocks (HSBs) and two highly variable blocks of insertions/deletions (INDELs). Most of the length variation among species occurred as varying numbers of nucleotides in the two INDELs. Little or no intraspecific sequence variation was detected over spatial scales of up to approximately 150 km. Interspecific sequence variation was lowest among the octocorals and greatest among the ceriantipatharians. Our data indicate that the orders Ceriantharia and Antipatharia are highly divergent, and a phylogenetic reconstruction provides support for the two-subclass system of the class Anthozoa (Hexacorallia and Octocorallia).</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"5 1","pages":"15-28"},"PeriodicalIF":0.0,"publicationDate":"1996-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19835019","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Do dinoflagellates contain a Cdc2-like protein kinase?","authors":"P Salois,&nbsp;D Morse","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>A protein present in extracts of the dinoflagellate Gonyaulax that was capable of binding an antibody directed against the conserved Cdc2 kinase epitope EGVPSTAIREISLLKE was characterized by Western blot analysis and DNA sequencing and shown not to encode a Cdc2 kinase. The amount, size, and isoelectric point of the immunoreactive species were invariant over a 24-hour period (encompassing S and M phases), and the DNA sequence of a cDNA isolated by immunologic screening showed that no conserved kinase regions were present in the deduced amino acid sequence. A method based on polymerase chain reaction (PCR), using primers designed from conserved regions in the Cdc2 kinases, was also unsuccessful in isolating a cdc2 gene homologue, although other kinases were identified.</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"5 1","pages":"52-61"},"PeriodicalIF":0.0,"publicationDate":"1996-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19835021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Growth enhancement in transgenic tilapia by ectopic expression of tilapia growth hormone.","authors":"R Martínez,&nbsp;M P Estrada,&nbsp;J Berlanga,&nbsp;I Guillén,&nbsp;O Hernández,&nbsp;E Cabrera,&nbsp;R Pimentel,&nbsp;R Morales,&nbsp;F Herrera,&nbsp;A Morales,&nbsp;J C Piña,&nbsp;Z Abad,&nbsp;V Sánchez,&nbsp;P Melamed,&nbsp;R Lleonart,&nbsp;J de la Fuente","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The generation of transgenic fish with the transfer of growth hormone (GH) genes has opened new possibilities for the manipulation of growth in economically important fish species. The tilapia growth hormone (tiGH) cDNA was linked to the human cytomegalovirus (CMV) enhancer-promoter and used to generate transgenic tilapia by microinjection into one-cell embryos. Five transgenic tilapia were obtained from 40 injected embryos. A transgenic animal containing one copy of the transgene per cell was selected to establish a transgenic line. The transgene was stably transmitted to F1 and F2 generations in a Mendelian fashion. Ectopic, low-level expression of tiGH was detected in gonad and muscle cells of F1 transgenic tilapia by immunohystochemical analysis of tissue sections. Nine-month-old transgenic F1 progeny were 82% larger than nontransgenic fish at p = .001. These results showed that low-level ectopic expression of tiGH resulted in a growth acceleration in transgenic tilapia. Tilapia GH gene transfer is an alternative for growth acceleration in tilapia.</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"5 1","pages":"62-70"},"PeriodicalIF":0.0,"publicationDate":"1996-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19835022","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The cDNA sequence of the lactate dehydrogenase-A of the spiny dogfish (Squalus acanthias): corrections to the amino acid sequence and an analysis of the phylogeny of vertebrate lactate dehydrogenases.","authors":"D W Stock,&nbsp;D A Powers","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The cDNA sequence of the lactate dehydrogenase-A (LDH-A) of the spiny dogfish was determined. The deduced amino acid sequence differed from a previously determined protein sequence by 5%. Separate maximum parsimony analyses of the two sequences along with LDHs of other vertebrates resulted in shorter trees with the sequence presented here, as well as fewer equally parsimonious trees. The new sequence also indicates a greater conservation of length among vertebrate LDHs than was previously suspected. Analyses of the phylogeny of vertebrate LDHs resulted in a monophyletic grouping of LDH-As, from within which mammalian LDH-C is derived. The phylogeny of LDH-As did not exactly match the phylogeny of the organisms, raising the possibility of multiple origins and losses of a muscle-predominant gene. LDH-Bs appear to have shared a single origin.</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"4 4","pages":"284-94"},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19524465","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Wresting the muscle from mussel beards: research and applications.","authors":"L M Rzepecki,&nbsp;J H Waite","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Marine and zebra mussels secrete byssal beards to attach themselves opportunistically to hard surfaces in their environment. By doing this, they naturally earn a reputation as fouling pests. The protein precursors of byssus in mussels are being investigated in the hope not only of discovering specific measures against these marine foulers, but also to gain some insights into the technically challenging task of engineering adhesive bonds underwater. Although byssal proteins are all part of the bearded glue that bonds them to a surface, they can be subdivided into three types depending on the function that they serve in byssal threads: (1) fibrous proteins form the load-bearing cables in the core of the threads, (2) cuticular proteins form a protective coat around the cables, and (3) adhesive proteins connect the cables to a foreign surface. A flaw in any one of these will undermine a mussel's ability to attach. The fibrous proteins can be collagenous, silk-like, elastic, or any combination of these. Covering these are the cuticular proteins, which are distinguished by their surface coupling properties, tandemly repeated primary sequence, and their high content of lysine and the exotic amino acid 3,4-dihydroxyphenyl-L-alanine (DOPA). The adhesive proteins are of low molecular weight, contain DOPA, and assemble to form microcellular solids (foams). Several of these proteins are already attracting biotechnological attention as cell and tissue attachment factors, anticorrosives, and metal-sequestering reagents.</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"4 4","pages":"313-22"},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19524467","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid detection of Vibrio cholerae contamination of seafood by polymerase chain reaction.","authors":"I Karunasagar,&nbsp;G Sugumar,&nbsp;I Karunasagar,&nbsp;A Reilley","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The possibility of detecting Vibrio cholerae contamination of seafood using a technique based on polymerase chain reaction (PCR) was studied. Direct PCR on lysate prepared from fish homogenates containing 10(3) V. cholerae/ml gave a positive reaction. When combined with alkaline peptone water (APW) enrichment, homogenates containing 1.4 cells/ml gave amplification signal. The technique could also detect V. cholerae O139, the recent epidemic serotype in the Indian subcontinent. An environmental isolate of non-O1 V. cholerae that produced cholera toxin was also positive in this assay. The results suggest that PCR-based techniques have great potential in quick detection of toxigenic V. cholerae in seafoods.</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"4 4","pages":"365-8"},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19522223","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rhamnolipid biosurfactant enhancement of hexadecane biodegradation by Pseudomonas aeruginosa.","authors":"G S Shreve,&nbsp;S Inguva,&nbsp;S Gunnam","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Mutants of Pseudomonas aeruginosa that produce and do not produce rhamnolipid biosurfactant are used to investigate the influence of cell-associated biosurfactant on cellular association with the hydrocarbon-water interface and on hydrocarbon uptake. Rhamnolipid-nonproducing mutant 65E12 of P. aeruginosa is unable to grow in minimal media containing hexadecane as a carbon source in the absence of exogenously added surfactant. Mutant PG201::rhlR grows very slowly in the absence of exogenously added surfactants. Both mutants are deficient in the positive regulatory gene controlling the activation of rhamnolipid synthesis. 65E12 is a double mutant that is also deficient in lipopolysaccharide synthesis. However, growth on hexadecane may be restored to varying degrees when small amounts of purified rhamnolipids or the synthetic anionic surfactant alkyl benzene sulfonate (ABS) is added to the cultures. Rhamnolipid biosurfactant is shown to be approximately 9 times more effective than the structurally similar synthetic anionic surfactant ABS in solubilizing hydrocarbon into the aqueous phase. Physical characteristics of the rhamnolipid and ABS micelles as determined by laser light scattering are described to explain the greater effectiveness of the rhamnolipid in solubilizing hexadecane. The cellular attachment to hydrocarbon-water interfaces and cellular aggregation of the wild-type and mutant strains are examined in the presence and absence of rhamnolipid or synthetic ABS surfactants. Differences in observed hexadecane degradation rates are explained on the basis of emulsified hexadecane concentration, cell surface hydrophobicity, and cellular localization in the culture.</p>","PeriodicalId":77273,"journal":{"name":"Molecular marine biology and biotechnology","volume":"4 4","pages":"331-7"},"PeriodicalIF":0.0,"publicationDate":"1995-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19522222","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}