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the midgut chymotrypsins of shrimps (penaeus monodon, penaeus japonicus and penaeus penicillatus).the midgut chymotrypsins (ec 3.4.4.5) of three species of shrimps, penaeus monodon, penaeus japonicus and penaeus penicillatus were purified and studied in detail to clarify previous ambiguity in their identification. in each of the species there are two major forms of chymotrypsin, both single-chained with three disulfide bonds. one has a pi of 3.2 and mr 27,000 or 28,000, while the other has a pi of 3.0 and mr 25,000 or 26,000. the n-terminal amino acid sequences of the p. monodon enzymes are ...19911657178
a heat-stable alkaline phosphatase from penaeus japonicus bate (crustacea: decapoda): a phosphatidylinositol-glycan anchored membrane protein.1. a heat-stable alkaline phosphatase was purified from penaeus japonicus, with a final specific activity of 21,280 u/mg of protein. 2. in polyacrylamide-gel electrophoresis under non-denaturing conditions, the purified shrimp alkaline phosphatase was found to have an identical molecular size and surface charge as the human placental enzyme. 3. by using sds-page, the monomers of shrimp alkaline phosphatase were discovered to have a mr 55,000 but those of human placental enzyme with a mr 70,000. ...19902331871
characterisation of shrimp haemocytes and plasma components by monoclonal antibodies.various haemolymph components of the shrimp penaeus japonicus were identified and characterised by monoclonal antibodies. three groups of monoclonal antibodies were raised. their reactivity to haemocyte types and/or secreted molecules was determined by immunofluorescence and the molecular masses of the antigens were analysed by western-blotting. a 170 kda protein, in reducing conditions, was recognized by four panhaemocytic monoclonal antibodies from group 1. this protein was present both in the ...19957622592
evolution of phosphagen kinase (iii). amino acid sequence of arginine kinase from the shrimp penaeus japonicus.the amino acid sequence of arginine kinase (ak) from the shrimp penaeus japonicus has been determined chemically. it consists of 355 amino acid residues, and has a calculated molecular mass of 40,018 da. the amino acid sequence of penaeus ak showed 91% and 51% identity, respectively, with those of aks from the lobster homarus vulgaris and the abalone nordotis madaka. it also showed significant homology (39-43%) with vertebrate or invertebrate creatine kinases and annelid glycocyamine kinase, sug ...19947765044
characterization of insulin receptor from the muscle of the shrimp penaeus japonicus (crustacea: decapoda).the beta-subunit of the insulin receptor from the muscle of the shrimp penaeus japonicus exists as multiple subtypes with m(r) of 79,000, 77,000 and 75,000. only the subunit of m(r) 79,000 is autophosphorylated after the addition of insulin. the autophosphorylation occurred specifically at tyr residues, as demonstrated by the specific subsequent dephosphorylation by the phosphotyrosyl protein phosphatase from the human placenta. the detergent, triton x-100, and the metal ion, mn2+, caused a noti ...19947881801
lipid and fatty acid composition of hepatopancreatic brush-border membrane vesicles from the prawn penaeus japonicus.brush-border membrane vesicles (bbmv) were isolated from prawn hepatopancreas by a procedure involving mg2+ preparation. the lipid composition of hepatopancreatic bbmv (hbbmv) was examined by the iatroscan tlc/fid technique, and the fatty acid profile of total lipids was determined by capillary gas chromatography. hbbmv were characterized by a high content of phospholipids and cholesterol. however, the cholesterol/phospholipid molar ratio is lower in prawn than in vertebrate bbmv, which agrees w ...19938340355
isolation of vibrio harveyi from diseased kuruma prawns penaeus japonicus.outbreaks of high mortality among the cultured kuruma prawn penaeus japonicus without overt gross signs occurred during august and december of 1994 in i-lan, taiwan. eleven luminous bacterial strains were isolated from the hepatopancreas of moribund prawns from five different farms by use of tryptic soy agar (tsa, supplemented with 2% nacl) and/or thiosulfate citrate bile salt sucrose (tcbs) agar. these strains, together with our two previously unpublished isolates, were characterized and identi ...19968662185
modification of dna topoisomerase i enzymatic activity with phosphotyrosyl protein phosphatase and alkaline phosphatase from the hepatopancreas of the shrimp penaeus japonicus (crustacea:decapoda).dna topoisomerase i was partially purified from the hepatopancreas of the shrimp penaeus japonicus. the specific activity of the final preparation was 7,000,000 units/mg of protein with sv40 viral dna as substrate. sdd-polyacrylamide gel electrophoresis of the final preparation yielded two major bands of proteins with m(r) 70,000 and m(r) 67,000, as well as less intense bands of proteins with m, 64,000 and m(r) 56,000. incubation of the partially purified enzyme fraction with rabbit antiserum ag ...19968759289
evidence for circulating hemocyte proliferation in the shrimp penaeus japonicus.we have observed by flow cytometric analysis that a small but consistent percentage of circulating hemocytes in phases s, g2 and m of the cell cycle can be detected in penaeus japonicus. significantly increased percentages of proliferating hemocytes (approximately three-fold increase) were observed after stimulation by lipopolysaccharide (lps), p43 (an immunosuppressive lymphocyte mitogenic protein produced by candida albicans) and a combination of lps and p43. moreover, an approximately six-fol ...19968799615
amino acid sequence of a peptide with molt-inhibiting activity from the kuruma prawn penaeus japonicus.six major peptides (pej-sgp-i-vi) that belong to the chh family have been isolated from the sinus gland extracts of the kuruma prawn penaeus japonicus. by in vitro assay using the y-organ of the crayfish procambarus clarkii, pej-sgp-iv was found to be active in inhibiting ecdysteroid synthesis. we determined the complete amino acid sequence. pej-sgp-iv consists of 77 amino acid residues, with both free aimno- and carboxyl-termini. the sequence of pej-sgp-iv shows considerable similarity to that ...19968801521
toxic effects of blooms of marine species of oscillatoriales on farmed prawns (penaeus monodon, penaeus japonicus) and brine shrimp (artemia salina).benthic and planktonic blooms of species of oscillatoriales coincided with mortalities of penaeus monodon during four episodes at australian prawn farms. oscillatoria corakiana was the dominant planktonic species at 65-90,000 cells/ml, but spirulina sp., lyngbya sp., oscillatoria sp. and nodularia sp. were also identified from the water column, benthic layers or surface mats. the levels and variety of vibrionaceae in prawn tissue, suggest that mortalities were caused by secondary infections of b ...19968875773
isolation and characterization of vibrio alginolyticus isolated from diseased kuruma prawn, penaeus japonicus.outbreaks of serious mortality among cultured kuruma prawns (penaeus japonicus) with white spotted syndrome in the carapace occurred in the summer of 1993 in i-lan, taiwan. a swarming bacterium, strain swy, was isolated from the hepatopancreas of the moribund prawns using tryptic soy agar supplemented with 1% nacl and/or thiosulphate citrate bile salt sucrose agar. this strain was characterized and identified as vibrio alginolyticus on the basis of a number of biochemical tests. the swy strain w ...19968936369
isolation of cdna encoding a putative chitinase precursor in the kuruma prawn penaeus japonicus.amino acid sequences of chitinases have been determined in insects, plants, yeast, and bacteria, but not in crustaceans. we searched for chitinase-encoding cdna in the kuruma prawn penaeus japonicus by polymerase chain reaction (pcr) amplification of hepatopancreas cdna using degenerate oligonucleotide primers derived from the two conserved regions of known chitinases. using a pcr product as a probe, a cdna clone was isolated. this clone contains an open reading frame for a protein (named pjchi- ...19968983198
alkaline serine protease is an exotoxin of vibrio alginolyticus in kuruma prawn, penaeus japonicus.an extracellular lethal toxin produced by vibrio alginolyticus strain swy originally isolated from diseased kuruma prawn(penaeus japonicus) was partially purified by fast protein liquid chromatography with hydrophobic interaction (phenyl sepharose hig hperformance) chromatography and gel filtration columns. the toxin is an alkaline serine protease, inhibited by phenyl-methylsulfonyl fluoride (pmsf),and showed maximal activity at ph 10, having a molecular weight of about 33kda estimated by sds-pa ...19979003588
cloning and sequence analysis of a cdna encoding a crustacean hyperglycemic hormone from the kuruma prawn penaeus japonicus.the crustacean hyperglycemic hormones (chhs) are released from the x-organ sinus gland complex and regulate the glucose level in the hemolymph. in the present study, we have isolated a complementary dna that encodes a chh (pej-sgp-iii) of the kuruma prawn penaeus japonicus to study its expression and the structure of its precursor. in a cdna clone of 774 base pairs (bp), an open reading frame of 345 bp was found. the conceptually translated protein consists of a signal peptide (24 residues), a c ...19979116871
amino acid sequences and activities of multiple hyperglycemic hormones from the kuruma prawn, penaeus japonicus.by means of a single-step reversed-phase hplc, six chh family peptides were isolated from sinus gland extracts of the kuruma prawn, penaeus japonicus. five of these peptides (pej-sgp-i, -ii, -iii, -v, and -vi) expressed hyperglycemic activity and were considered to be the hyperglycemic hormones of this prawn. the amino acid sequences of the five peptides were determined (the amino acid sequence of pej-sgp-iii had been previously determined), revealing that all consisted of 72 amino acid residues ...19979210164
effects of extracellular products of vibrio alginolyticus on penaeid prawn plasma components.the effects of both crude extracellular products (ecp) and a partially purified protease of vibrio alginolyticus on the plasma components of kuruma prawn (penaeus japonicus) and tiger prawn (p. monodon) were studied using crossed immunoelectrophoresis (cie). a component of the plasma, tentatively identified as coagulogen, apparently disappeared after incubation with the ecp, while the amount of a component tentatively identified as haemocyanin decreased. the coagulogen and an unknown component ( ...19979281857
molecular cloning of a molt-inhibiting hormone cdna from the kuruma prawn penaeus japonicus.the crustacean molt-inhibiting hormone (mih) is released from the x-organ sinus gland complex and suppresses ecdysteroid synthesis by the y-organ. in the present study, we have isolated a cdna which encodes a mih (pej-sgp-iv) of the kuruma prawn penaeus japonicus in order to study its expression and characterize the structure of its precursor. a cdna fragment was isolated using rt-pcr with two degenerate oligonucleotide primers that were designed based on the peptide sequence of pej-sgp-iv, and ...19979450390
chitinases are a multi-gene family in aedes, anopheles and drosophila.degenerate primers were used to amplify by the polymerase chain reaction (pcr) dna fragments from the chitinase genes of five insect species: aedes aegypti, anopheles freeborni, anopheles gambiae, anopheles stephensi and drosophila melanogaster. as many as four different products were found for each species; each deduced protein sequence having greatest homology to chitinase sequences from other species of insects and the crustacean, penaeus japonicus. the four pcr products of a. aegypti hybridi ...19989662472
cloning of a cdna encoding a putative molt-inhibiting hormone from the eyestalk of the sand shrimp metapenaeus ensis.degenerate primers were designed from the amino acid sequence of the neuropeptide pej-sgp-iv of the shrimp penaeus japonicus. reverse transcriptase-polymerase chain reaction (rt-pcr) was performed using eyestalk complementary dna of the sand shrimp metapenaeus ensis. a partial cdna that codes for a protein homologous to the neuropeptide pej-sg-iv was cloned. the partial cdna was used as a probe to screen the eyestalk cdna library. several cdna clones with nucleotide sequence identical to the par ...19989701616
a comparative study of three different isolates of white spot virus.three separate isolates of white spot virus (wsv) purified from 3 different penaeid shrimp species from different countries were compared morphologically, biochemically, and genomically using the following techniques; negative stain electron microscopy, sodium dodecyl-sulfate polyacrylamide gel electrophoresis/western blot, and restriction fragment length polymorphism (rflp), respectively. under the electron microscope, the 3 isolates were indistinguishable. their nucleoprotein cores exhibited t ...19989745720
[survey of disease of cultural shrimp in taiwan].the large number of hemocytes infiltrated several abnormal tissues of kuruma shrimp (penaeus japonicus), including musculature, hepatopancreas, lymphoid organ, gill filament and sponge tissue. in addition, there were many denatured hemocytes existing inside acidophilic particles and forming granules. futhermore, in hepatopancreas of kuruma shrimp, a white spot baculovirus (wsbv; 40-50 x 50-300 nm) was discovered in uh (undifferential haemocyte). the epithelium cells, which including stomach cuti ...19959774985
an infectious viral disease of penaeid shrimp newly found in korea.since 1993, massive mortalities have occurred among the penaeid shrimp penaeus orientalis cultured in korea. the major gross findings of the naturally occurring cases were 2 to 6 mm sized white spots on the inside of the carapace and reddish discolorization. amphophilic to basophilic intranuclear inclusion bodies were readily observed in the epithelia of epidermis, foregut, gills, and lymphoid organs. electron microscopy revealed enveloped and non-occluded ellipsoid to rod-shaped virus particles ...19989789980
mapping of dermatophagoides farinae mite allergens by two-dimensional immunoblotting.allergens from the house dust mite dermatophagoides farinae are responsible for frequent respiratory allergic disorders, but only 3 groups of these allergens are well characterized.19989802372
isolation of a cdna encoding a homologue of ribosomal protein l26 in the decapod crustacean penaeus japonicus.a complementary dna (pjl26) containing an open reading frame for a protein of 144 amino acids was isolated from the decapod crustacean penaeus japonicus. the conceptual pjl26 protein exhibits significant sequence similarities to ribosomal protein l26 in vertebrates. the level of the pjl26 messenger rna in the tail fan did not change significantly during the molt cycle. this cdna, therefore, may serve as a useful control probe in northern analyses of stage-specific transcripts in p. japonicus and ...19989892715
molecular cloning of ras cdna from penaeus japonicus (crustacea, decapoda): geranylgeranylation and guanine nucleotide binding.a cdna was isolated from the shrimp penaeus japonicus by homology cloning. the shrimp hepatopancreas cdna encodes a 187-residue polypeptide whose predicted amino acid sequence shares 85% homology with mammalian k-ras 4b protein and demonstrates identity in the guanine nucleotide binding domains. expression of the shrimp cdna in escherichia coli yielded a 21-kda polypeptide with a positive reactivity towards the monoclonal antibodies against mammalian ras. the gtp binding of the shrimp ras-encode ...19989931465
facilitated geranylgeranylation of shrimp ras-encoded p25 fusion protein by the binding with guanosine diphosphate.a cdna was isolated from the shrimp penaeus japonicus by homology cloning. similar to the mammalian ras proteins, this shrimp hepatopancreas cdna encodes a 187-residue polypeptide whose predicted amino acid sequence shares 85% homology with mammalian kb-ras proteins and demonstrates identity in the guanine nucleotide binding domains. expression of the cdna of shrimp in escherichia coli yielded a 25-kda polypeptide with positive reactivity toward the monoclonal antibodies against ras of mammals. ...199910194860
expression of a recombinant molt-inhibiting hormone of the kuruma prawn penaeus japonicus in escherichia coli.the crustacean molt-inhibiting hormone (mih) suppresses ecdysteroid synthesis by the y-organ. the mih of the kuruma prawn penaeus japonicus has recently been isolated and its cdna cloned. in this study, we expressed the mih in escherichia coli to obtain a large quantity of this hormone with biological activity. the mih cdna was processed and ligated into an expression plasmid. e. coli was transformed with this plasmid, and then the recombinant mih (r-mih) was expressed. the r-mih was put through ...199910540746
purification and partial characterization of a toxic serine protease produced by pathogenic vibrio alginolyticus.an extracellular lethal toxin produced by vibrio alginolyticus strain swy originally isolated from diseased kuruma prawn (penaeus japonicus) was purified using the akta purifier system with hydrophobic interaction chromatography, anion exchange and gel filtration columns. the toxin is an alkaline serine protease, inhibited by phenyl methylsulphonyl fluoride (pmsf), antipain and shows maximal activity at ph 8 to 11, having a pi of 4.3 and a molecular weight of approximately 33 kd. the toxin was c ...199910624008
primary culture of lymphoid organ cells and haemocytes of kuruma shrimp, penaeus japonicus.a primary cell culture system was developed for the cells of lymphoid organ tissue of kuruma shrimp, penaeus japonicus. minced tissues of lymphoid organs were seeded and incubated at 30 degrees c in medium 199 supplemented with 20% foetal bovine serum, a salt mixture and a lactalbumin hydrolysate (0.1 g/l). fibroblast-like cells and epithelioid-like cells survived for 54 days. cells did not survive after trypsin, collagenase or hyaluronidase treatment used for cell dissociation. mitogens (con a, ...199910627678
cloning and characterization of a novel nuclease from shrimp hepatopancreas, and prediction of its active site.approximately 95% of the amino acid sequence of a shrimp (penaeus japonicus) nuclease was derived from protease-digested peptides. a 1461-base cdna for the nuclease was amplified and sequenced with degenerate primers based on the amino acid sequence and then specific primers by 3' and 5' race (rapid amplification of cdna ends). it contains an open reading frame encoding a putative 21-residue signal peptide and a 381-residue mature protein. the n-terminus of the enzyme is pyroglutamate, deduced f ...200010698709
lethal attribute of serine protease secreted by vibrio alginolyticus strains in kuruma prawn penaeus japonicus.toxicity of the extracellular products (ecp) and the lethal attribute of serine protease secreted by five pathogenic vibrio alginolyticus strains from various sources in kuruma prawn penaeus japonicus were studied. the ecps of organisms originally isolated from diseased kuruma prawn or small abalone haliotis diversicolor supertexta were more lethal (ld50 value of 0.48 or 0.41 microg protein/g prawn) than those from diseased tiger prawn p. monodon, yellowfin porgy acanthopagrus latus or horse mac ...200010739107
detection of australian gill-associated virus (gav) and lymphoid organ virus (lov) of penaeus monodon by rt-nested pcr.a highly sensitive test based on reverse transcription followed by nested polymerase chain reaction (rt-npcr) was developed to detect the australian yellow-head-like viruses, gill-associated virus (gav) and lymphoid organ virus (lov) of penaeus monodon. the rt-npcr detected viral rna in as little as 10 fg lymphoid organ total rna isolated from gav-infected p. monodon. amplification of serial dilutions of a gav cdna clone showed that the nested pcr was sufficiently sensitive to detect a single ge ...200010768283
the penaeus monodon chitinase 1 gene is differentially expressed in the hepatopancreas during the molt cycle.we have isolated a full-length chitinase complementary dna from the tiger shrimp penaeus monodon that encodes a 621 amino acid protein possessing the functional domains of the chitinase protein family. the penaeus monodon chitinase 1 (pmchi-1) gene product is 81.8% identical to a chitinase 1 protein expressed in the hepatopancreas of penaeus japonicus. analysis by reverse transcription-polymerase chain reaction (rt-pcr) indicates that pmchi-1 messenger rna is detectable in the hepatopancreas and ...200010811951
molecular cloning of the crustacean dd4 cdna encoding a ca(2+)-binding protein.a cdna, named dd4, was identified in the prawn penaeus japonicus in a search for genes that were expressed during calcification of the crustacean exoskeleton. dd4 transcripts were detected in the epidermal cells underlying the exoskeleton specifically during the postmolt stage, when the calcification takes place. in the dd4 cdna an open reading frame of 542 amino acids was found. the deduced protein was acidic and proline-rich, and exhibited similarity to the drosophila ca(2+)-binding protein ca ...200011006119
enhancement of disease resistance against penaeid acute viraemia and induction of virus-inactivating activity in haemolymph of kuruma shrimp, penaeus japonicus, by oral administration of pantoea agglomerans lipopolysaccharide (lps). 200011016589
quasi-immune response of penaeus japonicus to penaeid rod-shaped dna virus (prdv).a quasi-immune response was demonstrated in kuruma prawn penaeus japonicus infected naturally or experimentally with prdv (penaeid rod-shaped dna virus, also called white spot syndrome virus or wssv), the causative agent of pav (penaeid acute viremia). in the first step of this study, natural survivors 4 mo after a pav outbreak demonstrated 94 % relative percent survival (rps) upon experimental prdv challenge. mortalities after challenge were confirmed by prdv detection to be due to pav using a ...200011023247
interaction of shrimp ras protein with mammalian caveolin-1.balb/3t3 cells were transformed by transfection with dna encoding the mutated ras(q(61)k) from shrimp penaeus japonicus (huang and chuang. 1999. j exp zool 283:510-521). the caveolin-1 in the membrane fraction extractable with 2% octyl glucoside was significant reduced, compared to untransformed cells. to understand this in more detail, the interaction of s-ras with caveolin was investigated using caveolin-1 purified from rat lungs. the purified caveolin-1 binds c-src, suppressing its autophosph ...200011074455
differences in the susceptibility of some penaeid prawn species to gill-associated virus (gav) infection.four species of penaeid prawn cultured in australia (penaeus monodon, penaeus esculentus, marsupenaeus japonicus and fenneropenaeus merguiensis) were injected with a virulent preparation of gill-associated virus (gav). p. monodon (average weight = 8.9, 13.9 and 19.2 g), p. esculentus (average weight = 19.5 g), f. merguiensis (average weight = 10.5 g), and small (average weight = 5.8 g) m. japonicus displayed overt signs of disease and mortalities which reached 82 to 100% within 23 d post-injecti ...200011104074
red swamp crawfish (procambarus clarkii): an alternative experimental host in the study of white spot syndrome virus.the pathogenicity of white spot syndrome virus (wssv) for the red swamp crawfish (procambarus clarkii) was investigated after infection by intramuscular (i.m.) injection and oral route. the cumulative mortality of crawfish injected i.m. with wssv reached 100% in 5 days. after oral feeding wssv-infected kuruma shrimp (penaeus japonicus) muscle tissues to the crawfish the cumulative mortality of this host reached 100% in 11 days. on reinfection trials, all the crawfish fed wssv-infected crawfish m ...200011332281
disrupting the transforming activity of shrimp ras(q(61)k) by deleting the caax box at the c-terminus.balb/3t3 cells were transformed by transfection with dna encoding the mutated ras(q(61)k) from shrimp penaeus japonicus. ras transcription and protein levels had increased significantly in the cells transfected with the s-ras plasmid, compared to cells transfected with a control plasmid pcdna3.1. the bacterially expressed gtp-locked s-ras(q(61)k) is successfully prenylated by rat protein geranylgeranyltransferase i (pggtase i) and then polymerized with tubulin, in agreement with findings for gtp ...200111351331
sequence analysis of a fish vitellogenin cdna with a large phosvitin domain.vitellogenins (vtg) are egg-yolk precursor proteins crucial for reproductive success in oviparous animals. we have cloned the first complete cichlid vtg cdna from the tilapia fish, oreochromis aureus. this cdna has the largest phosvitin (pv) domain amongst piscine vtgs, being comparable to those of lamprey, xenopus, and chicken. thus, the size of pv is independent of the evolutionary advancement of a species. the closer interspecific relationship between o. aureus vtg1 and fundulus vtgii than th ...200111602355
gtpase stimulation in shrimp ras(q(61)k) with geranylgeranyl pyrophosphate but not mammalian gap.balb/3t3 cells were transformed by transfection with dna encoding the mutated ras(q(61)k) from shrimp penaeus japonicus (huang et al., 2000). the gtpase-activating protein (gap) in the cytosol fraction was significantly expressed and degraded, compared to untransformed cells on the western blot. to understand this in more detail, the interaction of the bacterially expressed shrimp ras (s-ras) with gap was investigated using gap purified from mouse brains. sds-polyacrylamide gel electrophoresis r ...200111748613
effects of shrimp density on transmission of penaeid acute viremia in penaeus japonicus by cannibalism and the waterborne route.to investigate the effects of shrimp density on mortalities of penaeus japonicus in experimental penaeid acute viremia (= white spot syndrome), shrimp injected intramuscularly with penaeid rod-shaped dna virus (prdv) were reared at different densities. in expt 1, challenged (10(-6) dilution of a prdv preparation) shrimp were reared collectively in a tank or individually in separate chamber units. a significant difference in cumulative mortalities was found between collectively (75.6%) and indivi ...200111775794
identification and phylogeny of a non-specific endonuclease gene of white spot syndrome virus of shrimp.white spot syndrome virus (wssv) is a taxonomically unclassified virus which causes a disease in shrimps worldwide. a 936 bp long open reading frame (orf) was found on a 7.2 kb hindiii fragment of the dna genome of wssv located adjacent to the ribonucleotide reductase small subunit gene. this putative orf showed homology to prokaryotic and eukaryotic endonucleases, which contain a non-specific endonuclease motif. alignment with viral and eukaryotic endonuclease orfs revealed that most catalytica ...200111778701
selective enhanced phosphorylation of shrimp beta-tubulin by pkc-delta with pep(taxol), a synthetic peptide encoding the taxol binding region.beta-tubulin cdna from the shrimp penaeus japonicus was isolated by homology cloning. expression of cdna in escherichia coli yielded a 55 kda polypeptide, positive for monoclonal antibodies against mammalian beta-tubulin. autoradiography demonstrated the bacterially expressed hepatopancreas beta-tubulin of p. japonicus is specifically phosphorylated by the delta isoenzyme of protein kinase c (pkc-delta) purified from the plasma membrane of the shrimp heart, in the presence of the receptor for ac ...200211857471
gene expression in haemocytes of kuruma prawn, penaeus japonicus, in response to infection with wssv by est approach.gene expression in haemocytes of the kuruma prawn (penaeus japonicus) was investigated using an expressed sequence tag (est) approach. partial nucleotide sequences of cdna library clones constructed from normal and white spot syndrome virus (wssv)--infected p. japonicus haemocytes were determined. of 635 clones obtained from the normal library, 284 (44.7%) significantly matched sequences in genbank, and of 370 clones obtained from wssv-infected library, 174 (47.0%) significantly matched sequence ...200212201653
significance of a carboxyl-terminal amide moiety in the folding and biological activity of crustacean hyperglycemic hormone.recombinant peptides related to pej-sgp-i, one of several crustacean hyperglycemic hormones (chhs) existing in the kuruma prawn penaeus japonicus, were expressed in bacterial cells, and then purified after being allowed to refold. their circular dichroism spectra suggested that the recombinant pej-sgp-i having a free carboxyl-terminus (rpej-sgp-i-oh) differed slightly in secondary structure from the recombinant pej-sgp-i having an amidated c-terminus (rpej-sgp-i-amide). the hyperglycemic activit ...200212217413
molecular characterization of vitellin from the ovaries of the white shrimp penaeus (litopenaeus) vannamei.vitellin (vt) was purified from ovary extracts of mature females of the white shrimp penaeus vannamei using sepharose cl-4b and q-sepharose columns. native vt had an apparent molecular weight of 388 kda as detected in native-page, bound the lipophilic dye oil red o and had a total lipid content of approximately 43.8%. under reducing and denaturing conditions (sds-page), vt is composed of three major subunits of 87, 78 and 46 kda, although minor bands of 65, 61 and 31 kda are also detected. the 8 ...200212431404
genomic instability of prawn white spot bacilliform virus (wsbv) and its association to virus virulence.prawn white spot bacilliform virus (wsbv) is a major pathogen that causes prawn diseases. in this study, we examined the sequence of wsbv genome dna in the shrimp penaeus japonicus, p. vannamei, p. monodon, p. chinensis and metapenaeus ensis through successive pcr amplification of the dna fragments in the whole wsbv genome. we found a sequence deletion hotspot in the wsbv genome that is 305107 bp in length. the sizes of the deleted fragments were 4.6, 4.8 or 8.1 kbp depending on the species of p ...200212457981
a time-course study on the resistance of penaeus japonicus induced by artificial infection with white spot syndrome virus.the onset and duration of resistance in experimental survivors of penaeus japonicus produced by an intramuscular injection with white spot syndrome virus (wssv) were surveyed by re-challenge tests with the virus conducted at weeks 1-4 and months 1-3 post initial exposure (pie) to the virus. virus neutralising activity in the survivors' plasma was also examined. plasma-treated wssv was separated from the plasma by centrifugation and then injected into naïve shrimp, in parallel with each re-challe ...200212458745
effects of prenyl pyrophosphates on the binding of pkcgamma with rack1.receptors for activated c kinase (racks) are a group of pkc binding proteins that have been shown to mediate isoform-selective functions of pkc and to be crucial in the translocation and subsequent functioning of the pkc isoenzymes on activation. rack1 cdna from the shrimp penaeus japonicus was isolated by homology cloning. the hepatopancreas cdna from this shrimp was found to encode a 318-residue polypeptide whose predicted amino acid sequence shared 91% homology with human g(beta2)-like protei ...200312506405
[molecular cloning and sequence analysis of full length cdna encoding molt-inhibiting hormone from fennropenaeus chinensis].molt-inhibiting hormone (mih) is a neuropeptide member belonging to the eyestalk chh family. molting in shrimp is controlled by mih and ecdysone. by inhibiting the synthesis of ecdysone in the y-organ, mih indirectly suppress the molting activity of shrimp. a 697 bp full-length encoding molt-inhibiting hormone precursor cdna, which has been accepted by genbank (accession number: af469187), was firstly amplified from the total rna of eyestalk from fennropenaeus chinensis by the 3' and 5' rapid am ...200312776600
the effects of temperature on white spot syndrome infections in marsupenaeus japonicus. 200312877834
characterization and function of kuruma shrimp lysozyme possessing lytic activity against vibrio species.lysozyme cdna was isolated from a kuruma shrimp, marsupenaeus japonicus, hemocyte cdna library. the cdna consists of 1055 base pairs (bp) and encodes a chicken-type (c-type) lysozyme with a deduced amino acid sequence of 156 residues. the kuruma shrimp lysozyme has a high identity (79.7%) with pacific white shrimp lysozyme, and low to moderate identities (33.3-43.0%) with lysozymes of insects and vertebrates. comparisons with other c-type lysozymes from invertebrates and vertebrates showed that ...200314563565
ovarian primary tissue culture of the kuruma shrimp marsupenaeus japonicus.cell growth in ovarian primary culture of the kuruma shrimp, marsupenaeus japonicus, was examined under various culture conditions. the best growth of ovarian cells was obtained in a culture system consisting of double strength of leibovitz-15 medium supplemented with 10% fetal bovine serum, glucose (1 g/l), proline (0.1 g/l), tc-yeastolate (1 g/l), and lactalbumin hydrolysate (1 g/l). the cells survived in this medium at 25 degrees c for 45 d. the epithelial-like cells predominated in 10-d-old ...200314613332
replication of white spot syndrome virus in ovarian primary cultures from the kuruma shrimp, marsupenaeus japonicus.propagation of white spot syndrome virus (wssv) was investigated in primary ovarian cultures from the kuruma shrimp marsupenaeus japonicus. a wssv strain, purified by sucrose density gradient centrifugation, was inoculated into 10-day-old primary ovarian cultures. wssv infection induced marked cytopathic effect (cpe) on primary ovarian cells. initially, virus-infected cells began to shrink 72 h post-inoculation, followed by the rounding of most cells which detached finally from flask surface. el ...200414715311
identification of two distinct molt-inhibiting hormone-related peptides from the giant tiger prawn penaeus monodon.six peptides belonging to the crustacean hyperglycemic hormone (chh) family were isolated from extracts of the sinus glands of the giant tiger prawn penaeus monodon by reverse-phase high-performance liquid chromatography. these were designated pem-sgp-a to pem-sga-f (pem, penaeus monodon; sgp, sinus gland peptide) and their amino-terminal amino acid sequences were analyzed. five of the 6 peptides (pem-sgp-a, -b, -d, -e, and -f) exhibited similar amino acid sequences to those of chh peptides that ...200214961272
the first internal transcribed spacer (its-1) of ribosomal dna as a molecular marker for phylogenetic and population analyses in crustacea.the objective of the present study is to explore the feasibility of using the first internal transcribed spacer (its-1) of ribosomal dna as a molecular marker for studying the interspecific and intraspecific genetic variations among crustaceans. we designed primers that could amplify its-1 from a majority of taxonomic groups of crustaceans. the gene was found to exhibit a high degree of length polymorphism among different groups, ranging from 182 bp in the barnacle balanus amphitrite to approxim ...200114961351
apoptosis does not play an important role in the resistance of 'immune' penaeus japonicus against white spot syndrome virus.we previously demonstrated that kuruma shrimp, penaeus japonicus, exposed to white spot syndrome virus (wssv) became resistant ('immune' shrimp) to subsequent challenge with the virus. the present study investigated the role of apoptosis in the 'immune' shrimp during a secondary challenge with wssv. when naive kuruma shrimp were intramuscularly injected with wssv at a high or low dose, apoptosis was often detected by tunel assay in the lymphoid organ (lo), mainly in the early stage of the infect ...200414986935
detection of white spot syndrome virus from stomach tissue homogenate of the kuruma shrimp (penaeus japonicus) by reverse passive latex agglutination.a reversed passive latex agglutination (rpla) assay was developed for detecting the white spot syndrome virus (wssv), which was formally named as penaeid rod-shaped dna virus (prdv) in japan, from stomach tissue homogenate of the kuruma shrimp (penaeus japonicus). using high-density latex particles and specific polyclonal antibody, wssv was detectable after 4h incubation. the hemolymph, the stomach, and the gills were extracted from a shrimp that had been infected experimentally with wssv, the v ...200415109815
fusarium incarnatum isolated from black tiger shrimp, penaeus monodon fabricius, with black gill disease cultured in vietnam.fusarium incarnatum was isolated from gill lesions of cultured black tiger shrimp, penaeus monodon, in every crop during 2000-2002 in nghe an province, vietnam. infected shrimps showed typical signs of black gill disease and mortalities about a month prior to harvest. detailed morphological examinations, as well as molecular phylogenic analyses based on partial nucleotide sequences of ribosomal dna, were made on the isolates. an artificial infection of kuruma prawn, penaeus japonicus, using two ...200415357709
differential gene expression profile in hepatopancreas of wssv-resistant shrimp (penaeus japonicus) by suppression subtractive hybridization.in order to find the immune-relevant factors responsible for the virus resistance in the wssv-resistant shrimp, a suppression subtractive hybridization method was employed to identify differentially expressed genes and their expression profile in the hepatopancreas of the virus-resistant penaeid shrimp. thirty five genes were identified from more than 400 clones, of which eight are found for the first time in penaeid shrimp. betagbp is the most abundant gene in our subtractive library except hem ...200515450750
phylogenetic relationships of 12 penaeoidea shrimp species deduced from mitochondrial dna sequences.dna sequences of an 847 bp fragment of mitochondrial cytochrome oxidase subunit i (coi) gene and a 514 bp fragment of 16s rrna gene were determined to examine the phylogenetic relationships of 12 penaeoidea shrimp species (penaeus chinensis, penaeus japonicus, penaeus penicillatus, penaeus vannamei, penaeus canaliculatus, trachypenaeus curvirostris, metapenaeus affinis, metapenaeus ensis, metapenaeopsis barbata, parapenaeus fissuroides, parapenaeopsis hardiwickii, solenocera crassicomis). both f ...200415524311
isolation and cdna cloning of ovarian cortical rod protein in kuruma prawn marsupenaeus japonicus (crustacea: decapoda: penaeidae).two cortical rod proteins having molecular weights of 28.6 kda and 30.5 kda were isolated from the mature ovary of marsupenaeus japonicus using gel filtration and reversed-phase hplc. analysis of the n-terminal amino acid sequence of the 28.6 kda molecule revealed that amino acid residues 1-21 corresponded to residues 9-29 of the 30.5 kda molecule. examination of homology using blast showed that 21 amino acids out of 29 residues of the 28.6 kda molecule, and 14 out of 29 residues of the 30.5 kda ...200415572862
cloning and characterization of a molt-inhibiting hormone-like peptide from the prawn marsupenaeus japonicus.recently, it was demonstrated by pcr amplification that an additional molt-inhibiting hormone (mih)-like peptide was present in the kuruma prawn marsupenaeus japonicus. in this study, a cdna encoding this peptide designated pej-mih-b was cloned. the pej-mih-b gene was expressed strongly in the nerve cord, and weakly in the eyestalk. it was possible to isolate pej-mih-b from the sinus glands in the eyestalks. the recombinant pej-mih-b expressed in escherichia coli showed low molt-inhibiting activ ...200515629537
detection of white spot syndrome virus (wssv) from hemolymph of penaeid shrimps penaeus japonicus by reverse passive latex agglutination assay using high-density latex particles.a simple reverse passive latex agglutination (rpla) method for detecting white spot syndrome virus (wssv) in the hemolymph of infected kuruma shrimp (penaeus japonicus) was developed. it was confirmed that wssv could be detected from the shrimp hemolymph when the latex particles blocked with a casein protein were used as detection reagent. it became clear from the result of the infection trial that viruses are detectable by rpla before the appearance of overt symptoms of this disease. in additio ...200415664062
molecular cloning and characterisation of prophenoloxidase from haemocytes of the white shrimp, litopenaeus vannamei.cdna encoding prophenoloxidase (propo) of the white shrimp litopenaeus vannamei was obtained from haemocytes by a reverse-transcription polymerase chain reaction (rt-pcr) and rapid amplification of cdna using oligonucleotide primers based on the propo sequence of tiger shrimp penaeus monodon, freshwater crayfish pacifastacus leniusculus, green tiger shrimp penaeus semisulcatus (accession no.: af521949) and kuruma shrimp marsupenaeus japonicus (accession no.: ab0733223). propo of l. vannamei was ...200515683918
p120-gap associated with syndecan-2 to function as an active switch signal for src upon transformation with oncogenic ras.balb/3t3 cells transfected with plasmids pcdna3.1-[s-ras(q(61)k)] of shrimp penaeus japonicus were applied to reveal a complex of p120-gap/syndecan-2 being highly expressed upon transformation. of interest, most of the p120-gap/syndecan-2 complex was localized at caveolae, a membrane microdomain enriched with caveolin-1. to confirm the molecular interaction between syndecan-2 and p120-gap, we further purified p120-gap protein from mouse brains by using an affinity column of hitrap-rack1 and expr ...200515752734
differential profile of genes expressed in hemocytes of white spot syndrome virus-resistant shrimp (penaeus japonicus) by combining suppression subtractive hybridization and differential hybridization.white spot syndrome virus (wssv) is the major viral pathogen of culture shrimp. although remarkable progress has been made in characterizing the wssv genome, information concerning the antiviral process of host is still limited. to identify the genes differentially expressed along with their expression profile in the hemocytes of the virus-resistant shrimp, suppression subtractive hybridization (ssh) and differential hybridization (dh) were employed. relying on the sequences identified in the su ...200515781131
in vivo effects of a recombinant molt-inhibiting hormone on molt interval and hemolymph ecdysteroid level in the kuruma prawn, marsupenaeus japonicus.in order to determine the function of molt-inhibiting hormone (mih) in vivo, we examined the effects of injecting of a recombinant mih on the molt interval and hemolymph ecdysteroid level in the kuruma prawn, marsupenaeus japonicus. the injection of recombinant mih significantly prolonged the molt interval (9.0 +/-0.4 days in the control group, 9.5+/-0.5 days in the 2500 ng/g-body weight/injection-group, mean+/-sd), and significantly decreased the hemolymph ecdysteroid level (ratio of levels bet ...200515795494
dimerize rack1 upon transformation with oncogenic ras.from our previous studies, we learned that syndecan-2/p120-gap complex provided docking site for src to prosecute tyrosine kinase activity upon transformation with oncogenic ras. and, rack1 protein was reactive with syndecan-2 to keep src inactivated, but not when ras was overexpressed. in the present study, we characterized the reaction between rack1 protein and ras. rack1 was isolated from balb/3t3 cells transfected with plasmids pcdna3.1-[s-ras(q61k)] of shrimp penaeus japonicus and rack1 was ...200515796907
detection of hepatopancreatic parvovirus (hpv) in wild shrimp from india by nested polymerase chain reaction (pcr).the prevalence of hepatopancreatic parvovirus (hpv) in wild penaeid shrimp samples from india was studied by nested polymerase chain reaction (pcr) using primers designed in our laboratory. the virus could be detected in 9 out of 119 samples by non-nested pcr. however, by nested pcr 69 out of 119 samples were positive. the pcr results were confirmed by hybridization with digoxigenin-labelled dna probe. shrimp species positive by non-nested pcr included penaeus monodon, penaeus indicus and penaeu ...200515819441
trap rack1 with ras to mobilize src signaling at syndecan-2/p120-gap upon transformation with oncogenic ras.hitrap-syndecan-2/p120-gap and hitrap-syndecan-2/rack1 affinity columns were applied to reveal that src tyrosine kinase was highly expressed in balb/3t3 cells transfected with plasmids pcdna3.1-[s-ras(q(61)k)] of shrimp penaeus japonicus. both columns were effective to isolate src tyrosine kinase. the selective molecular affinity for src was found to be stronger with hitrap-syndecan-2/rack1, as revealed with competitive rack1 to dislodge src from hitrap-syndecan-2/p120-gap. we thus challenged th ...200515823555
four differentially expressed cdnas in callinectes sapidus containing the rebers-riddiford consensus sequence.decapod crustaceans such as callinectes sapidus, the blue crab, provide unique opportunities to study proteins involved in biomineralization. subsequent to each molt, the previously deposited soft cuticle is calcified while the postecdysial layers are simultaneously deposited and mineralized. though the majority of the exoskeleton hardens, morphologically similar cuticle at the joints, called arthrodial membrane, remains flexible. it seems reasonable that hypodermal cells producing these cuticle ...200515939643
more than one type of transglutaminase in invertebrates? a second type of transglutaminase is involved in shrimp coagulation.coagulation (clot formation) forms a physical barrier to prevent the loss of body fluid and dissemination of microbes into the haemocoel after injury or infection. its quickness and efficiency are essential for the survival of invertebrates that rely solely on innate immunity. transglutaminase (tg) catalyses intermolecular or intramolecular epsilon-(gamma-glutamyl) lysine bond formation, resulting in a protein polymerisation, and plays a role in blood coagulation and post-translational protein r ...200515985293
species identification of five penaeid shrimps using pcr-rflp and sscp analyses of 16s ribosomal dna.dna-based molecular markers for differentiation of five penaeid shrimps (penaeus monodon, p. semisulcatus, feneropenaeus merguiensis, litopenaeus vannamei and marsupenaeus japonicus) were developed based on polymerase chain reaction-restriction fragment length polymorphism (pcr-rflp) and single-stranded conformation polymorphism (sscp) of 16s ribosomal (r) dna. differentiation of p. monodon, p. semisulcatus and l. vannamei can be unambiguously carried out by pcrrflp of 16s rdna(560) whereas p. s ...200516053718
physiological function and metabolism of free d-alanine in aquatic animals.aquatic crustaceans and some bivalve mollusks contain a large amount of free d-alanine (up to 100 mumol/g wet wt.) in their tissues. under high salinity stress, crustaceans and bivalve mollusks largely accumulate d- and l-alanine irrespective of species examined, together with l-glutamine, l-proline, and glycine of which increases are species dependent. these data indicate that d-alanine is one of the major compatible osmolytes responsible for the intracellular isosmotic regulation in the tissue ...200516141518
upregulation of actin-like gene expression in giant freshwater prawns macrobrachium rosenbergii infected with metschnikowia bicuspidata.the giant freshwater prawn macrobrachium rosenbergii is commercially cultured throughout the world including taiwan. from 1992 to 1995, taiwanese production decreased by approximately 50% due to disease. the yeast metschnikowia bicuspidata is considered to be one of the major causes of white muscle disease, but the molecular mechanism of its pathogenesis is not known. using rna differential display (dd) with muscle and hepatopancreatic tissue, we identified a 324 nucleotide (nt) message specific ...200516231645
development of gene transfer technology for black tiger shrimp, penaeus monodon.an effective foreign gene transfer method for shrimp would have several potential uses in the shrimp culture industry, such as in preventing infectious diseases. we evaluated two gene transfer methods and used black tiger shrimp, penaeus monodon, as a model target species. for a promoter, we used the 1,592-bp promoter region of the ef-1alpha gene, a house-keeping gene, of kuruma shrimp marsupenaeus japonicus. the promoter region was linked to either the gene for green fluorescence protein (gfp) ...200516254920
molecular cloning and characterisation of prophenoloxidase cdna from haemocytes of the giant freshwater prawn, macrobrachium rosenbergii, and its transcription in relation with the moult stage.expression of prophenoloxidase (propo) cdna was determined from haemocytes of the giant freshwater prawn macrobrachium rosenbergii by a reverse-transcription polymerase chain reaction (rt-pcr) and rapid amplification of cdna using oligonucleotide primers based on the propo sequence of tiger shrimp penaeus monodon, freshwater crayfish pacifastacus leniusculus, green tiger shrimp penaeus semisulcatus, kuruma shrimp marsupenaeus japonicus, and white shrimp litopenaeus vannamei. the propo of m. rose ...200616376571
studies on nitric oxide synthase activity in haemocytes of shrimps fenneropenaeus chinensis and marsupenaeus japonicus after white spot syndrome virus infection.the nitric oxide synthase (nos) activity in the haemocytes of shrimps fenneropenaeus chinensis (osbeck) and marsupenaeus japonicus (bate) was studied after white spot syndrome virus (wssv) infection to determine its characteristics in response to virus infection. first, the nos activity in haemocytes of shrimps was determined by the means of nbt reduction and changes in cell conformation. and the variations of nos activity in shrimps after challenge with wssv intramuscularly were evaluated throu ...200616403661
effect of sulfide on the immune response and susceptibility to vibrio alginolyticus in the kuruma shrimp marsupenaeus japonicus.kuruma shrimp marsupenaeus japonicus held in 34 per thousand seawater were injected with tryptic soy broth (tsb)-grown vibrio alginolyticus (2.7x10(6)cfu shrimp(-1)), and then placed in water containing concentrations of sulfide at 0 (control), 51, 106, 528 and 1050microgl(-1), respectively. after 12-144h, mortality of v. alginolyticus-injected shrimp exposed to 528 and 1102microgl(-1) sulfide was significantly higher than that of shrimp exposed to 51microgl(-1) sulfide and the control solution. ...200716713303
biochemical characterization and cloning of transglutaminases responsible for hemolymph clotting in penaeus monodon and marsupenaeus japonicus.to investigate the shrimp blood clotting enzyme, a transglutaminase in the hemocytes of penaeus monodon (abbreviated as tgh) was purified. tgh is an abundant homodimeric cytosolic protein with 84.2 kda subunits. it clotted shrimp plasma and incorporated fluorescent dansylcadaverine into succinyl casein upon activation by cacl(2) in vitro. ic(50) for the activation was 3 mm, which is below the shrimp plasma ca(2+) level. showing similar properties as other type ii transglutaminase, tgh was partic ...200616769260
cloning and characterisation of a prophenoloxidase from the haemocytes of mud crab scylla serrata.a prophenoloxidase (propo) cdna was cloned from the haemocytes of mud crab scylla serrata using oligonucleotide primers and rt-pcr. both 3'- and 5'-regions were isolated by rapid amplification of cdna end (race) method. analysis of the nucleotide sequence revealed that the cdna clone has a full length of 2663bp, with an open reading frame of 2019bp, a 124-bp 5'-untranslated region, and a 520-bp 3'-untranslated region containing a poly a signal. it encodes a protein of 673 amino acids with a pred ...200716806468
expression and characterisation of tiger shrimp penaeus monodon penaeidin (mo-penaeidin) in various tissues, during early embryonic development and moulting stages.a penaeidin family, mo-penaeidin was cloned from the haemocytes of tiger shrimp penaeus monodon using genomic polymerase chain reaction (pcr) by gene specific primers. analysis of nucleotide sequence revealed that this mo-penaeidin consists of 1348 bp containing one intron (680 bp) and two exons (210 and 458 bp). it has an open reading frame (orf) of 222 p, which encodes a protein of 74 amino acids including a signal peptide of 19 amino acids. the calculated molecular mass of the mature protein ...200716820207
purification and characterisation of a calcium-independent lectin (pjlec) from the haemolymph of the shrimp penaeus japonicus.a natural lectin (nominated pjlec) was isolated from haemolymph of the shrimp penaeus japonicus by affinity chromatography with fetuin-sepharose. the result of sds-page showed that the purified pjlec protein consisted of 37kda subunits. the native pjlec behaved as a 452kda protein in gel filtration chromatography. those data suggest that pjlec is composed of 12 subunits of similar molecular weight. pjlec has a broad spectrum of bacterial-agglutination activities against both gram-positive and gr ...200716824770
molecular cloning and characterisation of a proteinase inhibitor, alpha 2-macroglobulin (alpha2-m) from the haemocytes of tiger shrimp penaeus monodon.an alpha 2-macroglobulin (alpha2-m) gene was cloned from the haemocytes of tiger shrimp penaeus monodon by rt-pcr, cloning and sequencing of overlapping pcr and rapid amplification of cdna ends (race) method. analysis of the nucleotide sequence revealed that the alpha2-m cdna consists of 4876 bp with an open reading frame (orf) of 4494 bp, a 52 bp 5'-untranslated region, and a 327 bp 3'-untranslated region containing a poly a signal. the open reading frame encodes a protein of 1498 amino acids w ...200716982096
silencing shrimp white spot syndrome virus (wssv) genes by sirna.white spot syndrome virus (wssv) is a major shrimp pathogen causing large economic losses all over the world. so far, however, there is no efficient approach to control this virus. rna interference (rnai), which has been applied to silence virus genes in eukaryotic organisms. in this investigation, a specific 21bp short interfering rna (vp28-sirna) targeting a major envelope protein gene (vp28) of wssv was used to induce gene silencing in vivo in penaeus japonicus shrimp. it was found that the t ...200717011052
hepatopancreatic nuclease of black tiger shrimp penaeus monodon unlikely to be involved in viral triggered apoptosis.nucleases are phosphodiesterases that hydrolyze dna and/or rna. in a search for shrimp nucleases involved in apoptosis, we discovered a nuclease from hepatopancreatic cdna of the black tiger shrimp penaeus monodon. the full-length nuclease gene was amplified and revealed to contain 1668bp corresponding to 381 deduced amino acid residues in the mature enzyme. sequence analysis indicated 83% nucleic acid identity and 89% amino acid identity to a nuclease from the kuruma shrimp penaeus japonicus (a ...200717049271
purification of sinus gland peptides having vitellogenesis-inhibiting activity from the whiteleg shrimp litopenaeus vannamei.vitellogenesis-inhibiting hormone (vih) in crustacea belongs to the crustacean hyperglycemic hormone (chh)-family. to characterize multiple vih molecules in the whiteleg shrimp litopenaeus vannamei, seven chh-family peptides designated as liv-sgp-a, -b, -c, -d, -e, -f, and -g were purified by reversed-phase hplc and identified by n-terminal amino acid sequencing. the dose-response effects of these peptides on vitellogenin mrna levels were examined using in vitro incubation of ovarian fragments o ...200717357858
antilipopolysaccharide factor (alf) of mud crab scylla paramamosain: molecular cloning, genomic organization and the antimicrobial activity of its synthetic lps binding domain.antilipopolysaccharide factors (alfs) are small basic proteins that can bind and neutralize lipopolysaccharide (lps) and have broad spectrum antimicrobial activities. in this study, we describe the isolation of the full-length cdna encoding for alf peptide (alfsp) of mud crab, scylla paramamosain by sequencing a hemocyte cdna library and using the rapid amplification cdna end (race) method. a full-length alfsp cdna of 614 bp contains an open reading frame (orf) of 372 bp, encoding 123 amino acid ...200717368541
studies on the transmission of wssv (white spot syndrome virus) in juvenile marsupenaeus japonicus via marine microalgae.we studied the possible role that marine microalgae may play during the outbreaks of wss (white spot syndrome). in order to elucidate the possibility of marine microalgae carrying wssv (white spot syndrome virus), six marine microalgae (isochrysis galbana, skeletonema costatum, chlorella sp., heterosigma akashiwo, scrippsiella trochoidea, dunaliella salina) were co-cultured with adult marsupenaeus japonicus infected with wssv and were assayed daily by nested-pcr to study whether they could carry ...200717383676
bioassay evidence for the transmission of wssv by the harpacticoid copepod nitocra sp.white spot syndrome virus (wssv) is now one of the most devastating and virulent viral agents threatening the penaeid shrimp culture industry and has been responsible for serious economic losses for shrimp farms worldwide. one remarkable characteristic of wssv is its wide reservoir range, which contributes to its wide geographical distribution. among epizootiological surveys, there is substantial evidence for wssv-positive copepods found in shrimp farming ponds. therefore, copepods are suspected ...200817698077
molecular cloning and phylogenetic analysis on alpha2-macroglobulin (alpha2-m) of white shrimp litopenaeus vannamei.the full-length sequence of alpha2-macroglobulin (alpha2-m) was cloned from the hemocytes of white shrimp litopenaeus vannamei by reverse transcription polymerase chain reaction (rt-pcr), cloning and sequencing of overlapping pcr, and the rapid amplification of cdna ends method. the alpha2-m cdna consists of 4682bp with an open reading frame of 4479bp, a 52-bp 5'-untranslated region, and a 132-bp 3'-untranslated region. the predicted molecular mass of the mature protein (1475 amino acids) is 167 ...200817706773
requirement for shrimp caspase in apoptosis against virus infection.caspases are central effectors in apoptosis. in this investigation, a novel caspase gene (designated as pjcaspase) obtained from the marine shrimp marsupenaeus japonicus was found to be significantly upregulated in survivors of wssv-challenged shrimp, suggesting that it might be involved in shrimp antiviral immunity. as revealed by rnai assays, when the pjcaspase gene was silenced by gene-specific sirna, the wssv-induced apoptosis was significantly inhibited. the results showed that the pjcaspas ...200818068223
antiviral phagocytosis is regulated by a novel rab-dependent complex in shrimp penaeus japonicus.rab gtpases are involved in phagosome formation and maturation. however, the role of rab gtpases in phagocytosis against virus infection remains unknown. in this study, it was found that a rab gene ( pjrab) from marine shrimp was upregulated in virus-resistant shrimp, suggesting that rab gtpase was involved in the innate response to virus. the rnai and mrna assays revealed that the pjrab protein could regulate shrimp hemocytic phagocytosis through a protein complex consisting of the pjrab, beta- ...200818072731
identification of cdna encoding toll receptor, mjtoll gene from kuruma shrimp, marsupenaeus japonicus.toll receptors are cell-surface receptors acting as pattern recognition receptors (prrs) that are involved in the signaling pathway for innate immunity activation and are genetically conserved from insects to mammals. tolls from penaeid shrimp are found in white leg shrimp litopenaeus vannamei (ltoll) and black tiger shrimp penaeus monodon (pmtoll). however, the molecular ligand-recognition patterns and identification of these penaeid toll classes remain unknown. here, we report cdna cloning of ...200818191582
difference between hemocyanin subunits from shrimp penaeus japonicus in anti-wssv defense.hemocyanin, the respiratory protein of arthropods and mollusks, was found to function against bacterial and fungal invasion. in this study, we show that this protein complex could delay the infection of white spot syndrome virus (wssv) in vivo and cloned two hemocyanin subunits genes (pjhcl and pjhcy) from the shrimp penaeus japonicus. further investigation of their transcriptional regulation during viral invasion showed that in contrast to subunit pjhcy, pjhcl expression could be strongly induc ...200818234332
identification of putative crustacean neuropeptides using in silico analyses of publicly accessible expressed sequence tags.the development of expressed sequence tags (ests) for crustacean cdna libraries and their deposition in publicly accessible databases has generated a rich resource for peptide discovery in this commercially and ecologically important arthropod subphylum. here, we have conducted in silico searches of these databases for unannotated ests encoding putative neuropeptide precursors using the blast program tblastn, and have predicted the mature forms of the peptides encoded by them. the primary strate ...200818321503
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