Characterization of eastern oyster (Crassostrea virginica Gmelin) chromosomes by fluorescence in situ hybridization with bacteriophage P1 clones

Chromosome identification is an essential step in genomic research, which so far has not been possible in oysters. We tested bacteriophage P1 clones for chromosomal identification in the eastern oyster Crassostrea virginica, using fluorescence in situ hybridization (FISH). P1 clones were labeled with digoxigenin-11-dUTP using nick translation. Hybridization was detected with fluorescein-isothiocyanate-labeled anti-digoxigenin antibodies and amplified with 2 layers of antibodies. Nine of the 21 P1 clones tested produced clear and consistent FISH signals when Cot-1 DNA was used as a blocking agent against repetitive sequences. Karyotypic analysis and cohybridization positively assigned the 9 P1 clones to 7 chromosomes. The remaining 3 chromosomes can be separated by size and arm ratio. Five of the 9 P1 clones were sequenced at both ends, providing sequence-tagged sites that can be used to integrate linkage and cytogenetic maps. One sequence is part of the bone morphogenetic protein type 1b receptor, a member of the transforming growth factor superfamily, and mapped to the telomeric region of the long arm of chromosome 2. This study shows that large-insert clones such as P1 are useful as chromosome-specific FISH probes and for gene mapping in oysters.

The manganese superoxide dismutase gene in bay scallop Argopecten irradians: Cloning, 3D modelling and mRNA expression

A novel manganese superoxide dismutase (MnSOD) was cloned from bay scallop Argopecten irradians by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA of MnSOD was of 1207 bp with a 678 bp open reading frame encoding 226 amino acids. The deduced amino acid sequence contained a putative signal peptide of 26 amino acids. Sequence comparison showed that the MnSOD of A. irradians shared high identity with MnSOD in invertebrates and vertebrates, such as MnSOD from abalone Haliotis discus discus (ABG88843) and frog Xenopus laevis (AAQ63483). Furthermore, the 3D structure of bay scallop MnSOD was predicted by SWISS-MODEL Protein Modelling Server and compared with those of other MnSODs. The overall structure of bay scallop MnSOD was similar to those of zebrafish Danio rerio, fruit fly Drosophila melanogaster, Chinese shrimp Fenneropenaeus chinensis, human Homo sapiens, and had the highest similarity to scallop Mizuhopecten yessoensis and abalone H. discus discus. A quantitative real-time PCR (qRT-PCR) assay was developed to detect the mRNA expression of MnSOD in different tissues and the temporal expression in haemocytes following challenge with the bacterium Vibrio anguillarum. A higher-level of mRNA expression of MnSOD was detected in gill and mantle. The expression of MnSOD reached the highest level at 3 h post-injection with V. anguillarum and then slightly recovered from 6 to 48 h. The results indicated that bay scallop MnSOD was a constitutive and inducible protein and thus could play an important role in the immune responses against V anguillarum infection. (c) 2008 Elsevier Ltd. All rights reserved.

Cloning and expression analysis of a HSP70 gene from Pacific abalone (Haliotis discus hannai)

Heat shock protein 70 (HSP70), the primary member of HSPs that are responsive of thermal stress, is found in all multicellular organisms and functions mostly as molecular chaperon. The inducible HSP70 cDNA cloned from Pacific abalone (Haliotis discus hannai) using rapid amplification of cDNA ends (RACE), was highly homologous to other HSP70 genes. The full-length cDNA of the Pacific abalone HSP70 was 2631 bp, consisting of a 5'-terminal untranslated region (UTR) of 90 bp, a 3'-terminal UTR of 573 by with a canonical polyadenylation signal sequence AATAAA and a poly (A) tail, and an open reading frame of 1968 bp. The HSP70 cDNA encoded a polypeptide of 655 amino acids with an ATPase domain of 382 amino acids, the substrate peptide binding domain of 161 amino acids and a C-terminus domain of 112 amino acids. The temporal expression of HSP70 was measured by semi-quantitative RT-PCR after heat shock and bacterial challenge. Challenge of Pacific abalone with heat shock or the pathogenic bacteria Vibrio anguillarum resulted in a dramatic increase in the expression of HSP70 mRNA level in muscle, followed by a recovery to normal level after 96 h. Unlike the muscle, the levels of HSP70 expression in gills reached the top at 12 h and maintained a relatively high level compared with the control after thermal and bacterial challenge. The upregulated mRNA expression of HSP70 in the abalone following heat shock and infection response indicates that the HSP70 gene is inducible and involved in immune response. (c) 2006 Elsevier Ltd. All rights reserved.

黄土-红粘土化学风化与晚新生代气候变化——甘肃平凉白水剖面为例

Late Cenozoic has witnessed a series of climate-environmental change which ends with a transitional shift from greenhouse to icehouse conditions. In last two decades, scientists began to employ the tectonic uplift and its weathering effect to interpret the climatic changes during the late Cenozoic. However, this endeavor has partly been restricted by the lacking of regional and global chemical weathering data. The loess-red clay deposit in the Loess Plateau may record the weathering features of the detritus material from the wide range upwind of the Loess Plateau. Therefore geochemistry of the loess-plaeosol and red clay sequences may provide insights into the regional chemical weathering regime and the connection between the chemical weathering and the late Cenozoic climate-environmental change Here we selected 319 samples from the Baishui section near the Pingliang City, Gansu Province, and analyzed them with X-ray fluorescence. Based on the result, we reconstruct the chemical weathering history of the Baishui section since 6Ma. We chose CIA as the proxy for chemical weathering intensity. The CIA ratio in soil units is higher than in adjacent loess horizons, but lower than in the red clay, in good agreement with the field observation. The CIA ratios of the Baishui section correlates well with the global ice volume fluctuations, indicating that the global cooling may contribute a lot to the chemical weathering variations in Chinese Loess Plateau. There are at least 3 million-year time scale variations that can be identified in the chemical weathering intensity curve, i.e., between 3.3 to 2.1 Ma, 1.7 to 0.9 Ma and from 0.9Ma. We think these may reflect the combined effect of the tectonic uplift and ice sheets on monsoon intensity. Other time scale variations can be also observed. In the period between 2.4 and 0.8 Ma, the CIA record display the 400,000 years cycle, which may be resulted from the Tibetan uplift during the Pliocene-early Pleistocene which have significantly amplified the monsoon response sensitivity to the orbital-scale variations in insolation. From 1.2 Ma, the 100,000 years period became intensifying, and particularly after 0.8 Ma, the earlier monsoon response at 400,000 year periodicity was overwhelmed by the ice sheet forcing at 100,000 year periodicity. These may indicate that the expansions of the Northern Hemisphere ice sheets may have crossed a threshold, which enforce the monsoon responding at the 100,000 year periodicity.

东营凹陷下第三系岩性油气藏形成条件与预测

The Dongying depression, located in the northern part of the jiyang Sag in the Buohaiwan Basin, comprises one of the major oil-producing bases of the Shengli oil-field. The prediction and exploration of subtle or litho1ogical oil traps in the oil-field has become the major confronted target. This is also one of the frontier study areas in the highly-explored oil-bearing basins in East China and abroad. Based on the integrated analysis of the geological, seismic and logging data and the theories of sequence stratigraphy, tectono-stratigraphy and petroleum system, the paper has attempted to document the characteristics of the sequence stratigraphic and structural frameworks of the low Tertiary, the syndepositional faults and their control on deposition, and then to investigate the forming conditions and distribution of the tithological oil traps in the depression. The study has set up a set of analysis methods, which can be used to effectively analysis the sequence stratigraphy of inland basins and predict the distribution of sandstone reservoirs in the basins. The major achievements of the study are as follows: 1. The low Tertiary can be divided into 4 second-order sequences and 13 third-order sequences, and the systems tracts in the third-order sequences have been also identified based on the examination and correction of well logging data and seismic profiles. At the same time, the parasequences and their stacking pattern in the deltaic systems of the third member of the Shahejie Formation have been recognized in the key study area. It has been documented that the genetic relation of different order sequences to tectonic, climatic and sediment supply changes. The study suggested that the formation of the second-order sequences was related to multiple rifting, while the activity of the syndepositional faults controlled the stacking pattern of parasequences of the axial deltaic system in the depression. 2. A number of depositional facies have been recognized in the low Tertiary on the basis of seismic facies and well logging analysis. They include alluvial fan, fan delta or braided delta, axial delta, lowstand fan, lacustrine and gravity flow deposits. The lacustrine lowstand fan deposits are firstly recognized in the depression, and their facies architecture and distribution have been investigated. The study has shown that the lowstand fan deposits are the important sandstone reservoirs as lithological oil traps in the depression. 3. The mapping of depositional systems within sequences has revealed the time and special distrbution of depositional systems developed in the basin. It is pointed out that major elastic systems comprise the northern marginal depositional systems consisting of alluvial fan, fan delta and offshore lowstand fan deposits, the southern gentle slope elastic deposits composed of shallow lacustrine, braided delta and lowstand fan deposits and the axial deltaic systems including those from eastern and western ends of the depression. 4. The genetic relationship between the syndepositional faults and the distribution of sandstones has been studied in the paper, upper on the analysis of structural framework and syndepositional fault systems in the depression. The concept of structural slope-break has been firstly introduced into the study and the role of syndepositional faults controlling the development of sequence architecture and distribution of sandstones along the hinged and faulted margins have been widely investigated. It is suggested that structural styles of the structural slope-break controlled the distribution of lowstand fan deposits and formed a favorable zone for the formation of lithological or structure-lithological oil traps in the basin. 5. The paper has made a deep investigation into the forming condition and processes of the lithological traps in the depression, based the analysis of composition of reservoir, seal and resource rocks. It is pointed out that there were two major oil pool-forming periods, namely the end of the Dongying and Guangtao periods, and the later one is the most important. 6. The study has finally predicted a number of favorable targets for exploration of lithologieal traps in the depression. Most of them have been drilled and made great succeed with new discovered thousands tons of raw oil reserves.

金川二矿区水平矿层灾变失稳问题研究

This dissertation is mainly composed of seven chapters. Specifically, a preface which introduces the background and significance of the dissertation, the present state of study relating to the dissertation, the methods and frame of the study in the dissertation is presented in the first chapter. The second chapter is mainly focused on the present theories and methods about the study on the Problems of Catastrophic Destabilization which are induced by deep mining. In the third chapter, basic conditions of Jinchuan Deposit II are introduced, which include regional geological background, engineering geological conditions, in-situ stress, hydrological geological conditions, mining methods and the present state of the horizontal ole layer. The fourth chapter analysizes the problem of Catastrophic Destabilization of the horizontal ole layer with theoretical methods and gives an analytical solution of elastic foundation beam which two ends are fixed and a catastrophic model of the horizontal ole layer. With FLAC3D, the Problem of Catastrophic Destabilization is analyzed, the variation of displacement and stress of the horizontal ole layer is described, and a development of plastic zone in ore layer is given out. The contents in the sixth chapter is a preliminary design of monitoring system for the horizontal ole layer. At the end of the text, several comprehensive conclusions are given in the seventh chapter.