148 resultados para Blotting, Northern


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Dry mass, nitrogen and phosphorus content in belowground litter of four emergent macrophytes (Typha glauca Godr., Phragmites australis (Cav.) Trin., Scolochloa festucacea (Willd.) Link and Scirpus lacustris L.) were followed for 1.2 years in a series of experimental marshes, Delta Marsh, Manitoba. Litter bags containing roots and rhizome materials of each species were buried in unflooded soil, or soil flooded at three water depths (1–30, 31–60, > 60 cm). There were few differences in dry mass loss in unflooded or flooded soils, and depth of flooding also had little effect on decomposition rates. In the flooded sites, Scolochloa and Phragmites roots lost more mass (48.9–63.8% and 59.2–85.5%, respectively) after 112 days than Typha and Scirpus (36.3–43.6 and 37.0–47.2%, respectively). These differences continued through to the end of the study, except in the shallow sites where Scirpus roots lost more mass and had comparable mass remaining as Scolochloa and Phragmites. In the unflooded sites, there was little difference between species. All litters lost nitrogen (22.9–90.0%) and phosphorus (46.3–92.7%) during the first 112 days, then levels tended to remain constant. Decay rates for our belowground root and rhizome litters were comparable to published literature values for aboveground shoot litter of the same species, except for Phragmites roots and rhizomes which decomposed at a faster rate (−k = 0.0014−0.0032) than shoots (−k = 0.0003−0.0007, [van der Valk, A.G., Rhymer, J.M., Murkin, H.R., 1991. Flooding and the decomposition of litter of four emergent plant species in a prairie wetland. Wetlands 11, 1–16]).

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The dichloromethane dehalogenase gene (dcm) of Methylophilus was cloned into the plasmid pKYLX71-35S2 and fused with the 35S2 promotor,then transformed Arabidopsis by the infiltration mediated with Agrobacterium tumefaciens. Homozygous dcm seeds were obtained after several generations selection on the medium with 50 mg L -1 kanamycin.Northern blotting showed dcm mRNA was high in cytoplasm and the detection of DCM enzyme indicated that the lines containing high dcm mRNA expressed high DCM enzyme activities. Fig 4, Ref 9

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In this paper, taking the northern region of Changxing County for example, with ammonia nitrogen as a pollution assessment index, we used an improved export coefficient method for estimate polluting load of non-point source pollution (NSP) and the social pollution survey data in the study area to estimate point source pollution. By comparing the total pollution output and the national surface water environmental quality standards find that the whole study area achieves the second water quality standard. However, Jiapu Township exceeds the water quality standards seriously because of the superfluous point source pollution. The water quality of other Townships is good. Further analysis showed that different types of land use and proportions in the northern region of Changxing County have a significant impact on the non-point source pollution, the general law is farmland contributes the largest share of the non-point source pollution output, followed by residential area and bare land, besides, with the increase in the proportion of forest and the decrease of farmland and residential area, the non-point source pollution reduces gradually. © 2010 IEEE.

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A full-length Cks1 homologue gene, AmphiCks1, was identified in amphioxus, Branchiostoma belcheri tsingtauense. Sequence characteristics, phylogeny and patterns of expression during embryonic and larval development were established. The protein predicted from AmphiCks1 showed high sequence identity with vertebrate and invertebrate homologues. Protein structural studies and phylogenetic analysis suggested that Cks homologues are evolutionarily conserved. The AmphiCks1 transcript was detected in most early developmental stages by northern blotting and whole-mount in situ hybridization, suggesting a role for the gene in cell division. (c) 2005 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.

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Radiolarian distribution in surface sediments of 104 stations from northern and central South China Sea show that the abundance and diversity of radiolarians increase with the water depth and are related to radiolarian concentrations from the water column, diminished terrigenous input, variability in calcareous shell content and the rate of silica and carbonate dissolution in the deep sea. According to the appearances of individual species in surface sediments at particular depths, seven faunal boundaries distribution are recognized at water depths of 100, 450, 650, 1000, 1200, 1400 and 2500 m. Four radiolarian assemblages in the sediments were identified by applying clustering procedures. Geographic distributions of these four assemblages coincide with present-day hydrologic features of the surface waters in this area.

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Eighteen isolates of the red algae Chondrus crispus were collected from Northern Atlantic sites, together with C. ocellatus, C. yendoi and C. pinnulatus from the North Pacific. The nuclear rDNA internal transcribed spacer (ITS) was sequenced and compared, spanning both the ITS regions and the 5.8S rRNA gene. Percentage of nucleotide variation for C. crispus ranged from 0.3% to 4.0%. Phylogenetic analyses were performed using maximum parsimony (MP), neighbor-joining (NJ) and minimum evolution methods. They showed that two main clades existed within the C. crispus samples examined and that suggested C. crispus had a single Atlantic origin. The clustering however did not follow the geographic origin. We hypothesized that the current distribution of C. crispus populations might be a result of three main factors: temperature boundaries, paleoclimate and paleoceanography. ITS data exhibited abundant molecular information not only for phylogeographical investigation but also for systematics studies.

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Lectin is regarded as a potential molecule involved in immune recognition and phagocytosis through opsonization in crustacean. Knowledge on lectin at molecular level would help us to understand its regulation mechanism in crustacean immune system. A novel C-type lectin gene (Fclectin) was cloned from hemocytes of Chinese shrimp Fenneropenaeus chinensis by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA consists of 1482 bp with an 861 bp open reading frame, encoding 287 amino acids. The deduced amino acid sequence contains a putative signal peptide of 19 amino acids. It also contains two carbohydrate recognition domains/C-type lectin-like domains (CRD1 and CRD2), which share 78% identity with each other. CRD1 and CRD2 showed 34% and 30% identity with that of mannose-binding lectin from Japanese lamprey (Lethenteron japonicum), respectively. Both CRD1 and CRD2 of Fclectin have I I amino acids residues, which are relatively invariant in animals' C-type lectin CRDs. Five residues at Ca2+ binding site I are conserved in Fclectin. The potential Ca2+/carbohydrate-binding (site 2) motif QPD, E, NP (Gln-Pro-Asp, Glu, Asn-Pro) presented in the two CRDs of Fclectin may support its ability to bind galactose-type sugars. It could be deduced that Fclectin is a member of C-type lectin superfamily. Transcripts of Fclectin were found only in hemocytes by Northern blotting and RNA in situ hybridization. The variation of mRNA transcription level in hemocytes during artificial infection with bacteria and white spot syndrome virus (WSSV) was quantitated by capillary electrophoresis after RT-PCR. An exploration of mRNA expression variation after LPS stimulation was carried out in primarily cultured hemocytes in vitro. Expression profiles of Fclectin gene were greatly modified after bacteria, LPS or WSSV challenge. The above-stated data can provide us clues to understand the probable role of C-type lectin in innate immunity of shrimp and would be helpful to shrimp disease control. (c) 2006 Elsevier Ltd. All rights reserved.

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Manganese superoxide dismutase (MnSOD) plays an important role in crustacean immune defense reaction by eliminating oxidative stress. Knowledge on MnSOD at molecular level allows us to understand its regulatory mechanism in crustacean immune system. A novel mitochondrial manganese superoxide dismutase (mMnSOD) was cloned from hepatopancreas of Chinese shrimp Fenneropenaeus chinensis by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA consists of 1185 bp with a 660 bp open reading frame, encoding 220 amino acids. The deduced amino acid sequence contains a putative signal peptide of 20 amino acids. Sequence comparison showed that the mMnSOD of F. chinensis shares 88% and 82% identity with that of giant freshwater prawn Macrobrachium rosenbergii and blue crab Callinectes sapidus, respectively. mMnSOD transcripts were detected in hepatopancreas, hemocytes, lymphoid organ, intestine, ovary, muscle and gill by Northern blotting. RT-PCR analysis indicated that mMnSOD showed different expression profiles in shrimp hemocytes and hepatopancreas after artificial infection with while spot syndrome virus (WSSV). In addition, a fusion protein containing mMnSOD was produced in vitro. LC-ESI-MS analysis showed that two peptide fragments (-GDVNTVISLAPALK- and -NVRPDYVNAIWK-) of the recombinant protein were identical to the corresponding sequence of M. rosenbergii mMnSOD, and the enzyme activity of the refolded recombinant protein was also measured. (c) 2006 Elsevier Ltd. All rights reserved.

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Antimicrobial peptides are important components of the host innate immune responses by exerting broad-spectrum microbicidal activity against pathogenic microbes. The first mollusk big defensin (designated AiBD) cDNA was cloned from bay scallop Argopecten irradians by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) techniques. The scallop AiBD consisted of 531 nucleotides with a canonical polyadenylation signal sequence AATAAA and a poly(A) tail, encoding a polypeptide of 122 amino acids. The high similarity of AiBD deduced amino acid sequence with big defensin from Tachypleus tridentatus and Branchiostoma belcheri tsingtaunese indicated that AiBD should be a member of big defensin family. The expression of AiBD in various tissues was measured by using Northern blotting analysis. mRNA transcripts of AiBD could be detected in haemocytes of unchallenged scallops. The temporal expression of AiBD in haemolymph after Vibrio anguilarum challenge was recorded by quantitative real time PCR. The relative expression level of AiBD in haemolymph was up-regulated evenly in the first 8 h, followed by a drastic increase, and increased 131.1-fold at 32 h post-injection. These results indicated that AiBD could be induced by bacterial challenge, and it should participate in the immune responses of A. irradians. Biological activity assay revealed that recombinant AiBD could inhibit the growth of both Gram-positive and Gram-negative bacteria, and also showed strong fungicidal activity towards the expression host. Recombinant expression of AiBD made it possible to further characterize its functions involved in immune responses, and also provided a potential therapeutic agent for disease control in aquaculture. (c) 2006 Elsevier Ltd. All rights reserved.

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A new member of antimicrobial protein genes of the Crustin family was cloned from haemocytes of the Chinese shrimp Fenneropenaeus chinensis by 3' and 5' RACE. The full-length cDNA of Crustin-like gene contains a 390 bp open reading frame, encoding 130 amino acids. The deduced peptide contains a putative signal peptide of 17 amino acids and mature peptide of 113 amino acids. The molecular mass of the deduced mature peptide is 12.3 ku. It is highly cationic with a theoretical isoelectric point of 8.5. The deduced amino acids sequence of this Crustin showed high homology with those of Penaeus (Litopenaeas) setferus. Northern blotting showed that the cloned Crustin gene was mainly expressed in haemocytes, gill, intestine, and RNA in situ hybridization indicated that the Crustin gene was constitutively expressed exclusively in haemocytes of these tissues. Capillary electrophoresis RT-PCR analysis showed that Crustin was up-regulated dramatically from 12 to 48 h after a brief decrease of mRNA during first 6 h in response to microbe infection. The level of Crustin mRNA began to restore at 72 h post-challenge. This indicated that Crustin gene might play an important role when shrimps are infected by bacterial pathogen.

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Quantitative studies on the evolution and dynamics of the deepwater area of Pearl River Mouth basin (PRMB) were carried out based on the latest geological and seismic data. The study area is generally in an extensional state during the Cenozoic. The major extension happened in the earlier syn-rift stages before 23 Ma and the extension after 23 Ma is negligible. Two rapid subsidence periods, 32-23 Ma and 5.3-2.6 Ma, are identified, which are related to the abrupt heat decay during margin breakup and the collision between the Philippine Sea plate and the Eurasian plate, respectively. The strongest crustal thinning in the Baiyun (sic) sag may trigger the syn-rift volcanism along the weak faulted belt around the sag. The Cenozoic tectonic evolution of the study area could be divided into five stages: rifting (similar to 50-40 Ma), rift-drift transition (similar to 40-32 Ma), early post-breakup (similar to 32-23 Ma), thermal subsidence (similar to 23-5.3 Ma) and neotectonic movement (similar to 5.3-0 Ma).