Molecular cloning and stress-dependent expression of a gene encoding Delta(12)-fatty acid desaturase in the Antarctic microalga Chlorella vulgaris NJ-7

The psychrotrophic Antarctic alga, Chlorella vulgaris NJ-7, grows under an extreme environment of low temperature and high salinity. In an effort to better understand the correlation between fatty acid metabolism and acclimation to Antarctic environment, we analyzed its fatty acid compositions. An extremely high amount of Delta(12) unsaturated fatty acids was identified which prompted us to speculate about the involvement of Delta(12) fatty acid desaturase in the process of acclimation. A full-length cDNA sequence, designated CvFAD2, was isolated from C. vulgaris NJ-7 via reverse transcription polymerase chain reaction (RT-PCR) and RACE methods. Sequence alignment and phylogenetic analysis showed that the gene was homologous to known microsomal Delta(12)-FADs with the conserved histidine motifs. Heterologous expression in yeast was used to confirm the regioselectivity and the function of CvFAD2. Linoleic acid (18:2), normally not present in wild-type yeast cells, was detected in transformants of CvFAD2. The induction of CvFAD2 at an mRNA level under cold stress and high salinity is detected by real-time PCR. The results showed that both temperature and salinity motivated the upregulation of CvFAD2 expression. The accumulation of CvFAD2 increased 2.2-fold at 15A degrees C and 3.9-fold at 4A degrees C compared to the alga at 25A degrees C. Meanwhile a 1.7- and 8.5-fold increase at 3 and 6% NaCl was detected. These data suggest that CvFAD2 is the enzyme responsible for the Delta(12) fatty acids desaturation involved in the adaption to cold and high salinity for Antarctic C. vugaris NJ-7.

Identification of an Atlantic salmon IFN multigene cluster encoding three IFN subtypes with very different expression properties

A cluster of 11 interferon (IFN) genes were identified in the Atlantic salmon genome linked to the growth hormone I gene. The genes encode three different IFN subtypes; IFNa (two genes), IFNb (four genes) and IFNc (five genes), which show 22-32% amino acid sequence identity. Expression of the fish IFNs were studied in head kidney, leukocytes or To cells after stimulation with the dsRNA poly I:C or the imidazoquinoline S-27609. In mammals, poly I:C induces IFN-beta through the RIG-I/MDA5 or the TLR3 pathway, both of which are dependent on NF-kappa B. In contrast, S-27609 induces mammalian IFN-alpha in plasmacytoid dendritic cells through the TLR7 pathway independent of NF-kappa B. The presence of an NF-kappa B site in their promoters and their strong up-regulation by poly I:C, suggest that salmon IFNa1/IFNa2 are induced through similar pathways as IFN-beta. In contrast, the apparent lack of NF-kappa B motif in the promoter and the strong upregulation by S-27609 in head kidney and leukocytes, suggest that IFNb genes are induced through a pathway similar to mammalian IFN-alpha. IFNc genes showed expression patterns different from both IFNa and IFNb. Taken together, salmon IFNa and IFNb are not orthologs of mammalian IFN-beta and IFN-alpha, respectively, but appear to utilize similar induction pathways. (C) 2008 Elsevier Ltd. All rights reserved.

Cloning, sequencing and expression analysis of cDNA encoding a constitutive heat shock protein 70 (HSC70) in Fenneropenaeus chinensis

The cDNA encoding hsc70 of Chinese shrimp Fenneropenaeus chinensis was cloned from hepatopancreas by RT-PCR based on its EST sequence. The full length cDNA of 2090 bp contained an open reading frame of 1956 nucleotides and partial 5'- and 3'-untranslated region(5'- and 3'-UTR). PCR amplification and sequencing analysis showed the existence of introns in the region of 1-547 bp, but they did not exist in the region of 548-2090 bp of hsc70 cDNA. When the deduced 652 amino acid sequence of HSC70 was compared with the members of HSP70 family from other organisms, the results showed 85.9% similarity with HSC71 from Oncorhynchus mykiss and HSC70 from Homo sapiens. It also exhibited 85.8% similarity with HSP70 from Mus musculu and 85.4% with HSC70 from Manduca sexta. Expression analysis showed that hsc70 mRNA was espressed constitutively in hepatopancreas, muscle, eyestalks, haemocytes, heart, ovary, intestine and gills in Fenneropenaeus chinensis. No difference could be detected on hsc70 mRNA level in muscle between heat-shocked and control animals.

Recombinant expression of rt-PA gene (encoding Reteplase) in gametophytes of the seaweed Laminaria japonica (Laminariales, Phaeophyta)

The life cycle of seaweed Laminaria japonica involves a generation alternation between diploid sporophyte and haploid gametophte. The expression of foreign genes in sporophte has been proved. In this research, the recombinant expression in gametophyte was investigated by particle bombardment with the rt-PA gene encoding the recombinant human tissue-type plasminogen activator (Reteplase), which is a thrombolytic agent for acute myocardial infarction (AMI). Transgenic gametophytes were selected by their resistance to herbicide phosphiothricin (PPT), and proliferated in an established bubble column photo-bioreactor. According to the results from quantitative ELISA, Southern blotting, and fibrin agarose plate assay (FAPA) for bioactivity, it was showed that the rt-PA gene had been integrated into the genome of gametophytes of L. japonica, and the expression product showed the expected bioactivity, implying the proper post-transcript modification in haploid gametophyte.

There are two 5 '-flanking regions of bkt encoding beta-carotene ketolase in Haematococcus pluvialis

The unicellular green alga Haematococcus pluvialis accumulates a commercially valuable astaxanthin, with levels reaching up to 4% dry weight under environmental stress. In recent years, much effort has been devoted to understanding the molecular mechanisms regulating astaxanthin biosynthetic pathways. Beta-carotene ketolase (bkt), with control being exhibited at the transcription level, plays an important role in astaxanthin biosynthesis by H. pluvialis. Here we demonstrate the presence of two separate 5'-flanking regions [1.5 kilobase (kb) and 2 kb] of bkt (bkt1 and bkt2) that possess regulatory elements similar to those of known stress-responsive genes in plants. Results of 5'-deletion constructs and transient beta-galactosidase expression assays demonstrate that there may be positive regulatory elements governing expression in the shorter promoter at -1060/-900 from the 1.5 kb 5' region, and in the longer promoter at -1838/-1219 and at -1046/ -734 from the 2 kb 5' region relative to each homologous ATG start codon. Furthermore, our present studies reveal that the first intron (+371/+497) downstream from the 1.5 kb 5' untranslated region of bkt1 may function as a negative regulatory element to regulate its own promoter.

Diversity, abundance and distribution of amoA-encoding archaea in deep-sea methane seep sediments of the Okhotsk Sea

The authors would like to thank Jin Sun, Jian Sun, Liangliang Kong, Nianshuang Wang, Chunhui Wang, Linbao Zhang and Ying Zhang for their assistance in the project. This work was supported by China Ocean Mineral Resources R&D Association grants DYXM-115-02-2-20 and DYXM-115-02-2-6, Hi-Tech Research and Development Program of China grant 2007AA091903, China National Natural Science Foundation grant 40576069, National Basic Research Program of China grant 2009CB219506 and the Fundamental Research Funds for the Central Universities of China grant 09CX05005A. M. G. K. was funded by incentive funds provided by the UofL-EVPR office and the US National Science Foundation (EF-0412129).

Diversity and spatial distribution of amoA-encoding archaea in the deep-sea sediments of the tropical West Pacific Continental Margin

The ecological characteristics of the deep-sea amoA-encoding archaea (AEA) are largely unsolved. Our aim was to study the diversity, structure and distribution of the AEA community in the sediments of the tropical West Pacific Continental Margin, to develop a general view of the AEA biogeography in the deep-sea extreme environment. Archaeal amoA clone libraries were constructed. Diverse and novel amoA sequences were identified, with the Bohol Sea, Bashi Strait and Sibuyan Sea harbouring the highest and the Bicol Shelf the lowest AEA diversity. Phylogenetic and statistical analyses illustrate a heterogeneous distribution of the AEA community, probably caused by the differential distribution of the terrestrial or estuarine AEA in the various sampling sites. The deep-sea sedimentary environments potentially harbour diverse and novel AEA in the tropical West Pacific Continental Margin. The stations in the Philippine inland seas (including station 3043) may represent AEA assemblages with various terrestrial influences and the stations connected directly to the open Philippine Sea may represent marine environment-dominant AEA assemblages. Our study indicates the potential importance of geological and climatic events in the transport of terrestrial micro-organisms to the deep-sea sedimentary environments, almost totally neglected previously.

Diversity and Distribution of Sediment NirS-Encoding Bacterial Assemblages in Response to Environmental Gradients in the Eutrophied Jiaozhou Bay, China

A gene-clone-library-based molecular approach was used to study the nirS-encoding bacteria-environment relationship in the sediments of the eutrophic Jiaozhou Bay. Diverse nirS sequences were recovered and most of them were related to the marine cluster I group, ubiquitous in estuarine, coastal, and marine environments. Some NirS sequences were unique to the Jiaozhou Bay, such as the marine subcluster VIIg sequences. Most of the Jiaozhou Bay NirS sequences had their closest matches originally detected in estuarine and marine sediments, especially from the Chesapeake Bay, indicating similarity of the denitrifying bacterial communities in similar coastal environments in spite of geographical distance. Multivariate statistical analyses indicated that the spatial distribution of the nirS-encoding bacterial assemblages is highly correlated with environmental factors, such as sediment silt content, NH4+ concentration, and OrgC/OrgN. The nirS-encoding bacterial assemblages in the most hypernutrified stations could be easily distinguished from that of the least eutrophic station. For the first time, the sedimentological condition was found to influence the structure and distribution of the sediment denitrifying bacterial community.

High-power laser beam shaping by inseparable two-dimensional binary-phase gratings for surface modification of stamping dies

For surface modification of stamping dies, an inseparable two-dimensional binary-phase gratings is introduced to implement the wavefront transformation of high-power laser beams. The design and fabrication of the gratings are described in detail. Two-dimensional even sampling encoding scheme is adopted to overcome the limitations of conventional Dammann grating in the design of two-dimensional output patterns. High diffractive efficiency (>70%) can be achieved through the transformation of the Gaussian laser beam into several kinds of two-dimensional arrays in focal plan. The application of the binary-phase gratings in the laser surface modification of ductile iron is investigated, and the results show that the hardness and the wear resistance of the sample surface were improved significantly by using the binary-phase gratings. (C) 2008 Elsevier Ltd. All rights reserved.

Rotation Numbers Of Invariant Manifolds Around Unstable Periodic Orbits For The Diamagnetic Kepler Problem

In this paper, a method to construct topological template in terms of symbolic dynamics for the diamagnetic Kepler problem is proposed. To confirm the topological template, rotation numbers of invariant manifolds around unstable periodic orbits in a phase space are taken as an object of comparison. The rotation numbers are determined from the definition and connected with symbolic sequences encoding the periodic orbits in a reduced Poincare section. Only symbolic codes with inverse ordering in the forward mapping can contribute to the rotation of invariant manifolds around the periodic orbits. By using symbolic ordering, the reduced Poincare section is constricted along stable manifolds and a topological template, which preserves the ordering of forward sequences and can be used to extract the rotation numbers, is established. The rotation numbers computed from the topological template are the same as those computed from their original definition.

Repeat-until-success distributed quantum computation by using single-photon interference at a beam splitter

A repeat-until-success (RUS) measurement-based scheme for the implementation of the distributed quantum computation by using single-photon interference at a 50:50 beam splitter is proposed. It is shown that the 50:50 beam splitter can naturally project a suitably encoded matter-photon state to either a desired entangling gate-operated state of the matter qubits or to their initial state when the photon is detected. The recurrence of the initial state permits us to implement the desired entangling gate in a RUS way. To implement a distributed quantum computation we suggest an encoding method by means of the effect of dipole-induced transparency proposed recently [E. Waks and J. Vuckovic, Phys. Rev. Lett. 96, 153601 (2006)]. The effects of the unfavorable factors on our scheme are also discussed.

Parallel optical negabinary arithmetic based on logic operations

On the basis of signed-digit negabinary representation, parallel two-step addition and one-step subtraction can be performed for arbitrary-length negabinary operands.; The arithmetic is realized by signed logic operations and optically implemented by spatial encoding and decoding techniques. The proposed algorithm and optical system are simple, reliable, and practicable, and they have the property of parallel processing of two-dimensional data. This leads to an efficient design for the optical arithmetic and logic unit. (C) 1997 Optical Society of America.

Classified one-step modified signed-digit arithmetic and its optical implementation

Based on the two-step modified signed-digit (MSD) algorithm, we present a one-step algorithm for the parallel addition and subtraction of two MSD numbers. This algorithm is reached by classifying the three neighboring digit pairs into 10 groups and then making a decision on the groups. It has only a look-up truth table, and can be further formulated by eight computation rules. A joint spatial encoding technique is developed to represent both the input data and the computation rules. Furthermore, an optical correlation architecture is suggested to implement the MSD adder in parallel. An experimental demonstration is also given. (C) 1996 Society of Photo-Optical instrumentation Engineers.

Code conversion from signed-digit to complement representation based on look-ahead optical logic operations

We present, for the first time to our knowledge, a generalized lookahead logic algorithm for number conversion from signed-digit to complement representation. By properly encoding the signed-digits, all the operations are performed by binary logic, and unified logical expressions can be obtained for conversion from modified-signed-digit (MSD) to 2's complement, trinary signed-digit (TSD) to 3's complement, and quarternary signed-digit (QSD) to 4's complement. For optical implementation, a parallel logical array module using an electron-trapping device is employed and experimental results are shown. This optical module is suitable for implementing complex logic functions in the form of the sum of the product. The algorithm and architecture are compatible with a general-purpose optoelectronic computing system. (C) 2001 Society of Photo-Optical Instrumentation Engineers.

Encoded cell grating array in anti-counterfeit technology

The dot matrix hologram (DMH) has been widely used in anti-counterfeiting label. With the same technology and cell array configuration, we can encode to the incidence beam. These codes can be some image matrix grating with different grating gap and different grating orientation. When the multi-level phase diffractive grating is etched, the incidence beam on the cell appears as an encoding image. When the encoded grating and DMH are used in the same label synchronously, the technology of multi-encoded grating array enhances the anti-counterfeit ability.