Through data mining of the Cordyceps militaris genome, a lectin-encoding gene, CMLec3, was identified. In this study, the CMLec3 sequence was analyzed using bioinformatics approaches, and the gene was heterologously expressed in Escherichia coli BL21 cells. The biological activity of the product was examined. In addition, CMLec3 gene expression levels were assessed. The results showed that the CMLec3 protein contained a lectin domain structure and was successfully expressed. The CMLec3 protein partly inhibited HeLa cell proliferation. CMLec3 exhibited the highest gene expression in the primordium at a level 5.19 times that of the mycelium and 1.35 times that of the fruiting body. This suggests that the gene may be related to fruiting body development.

Basidiomycetes can degrade lignocellulosic biomass, and some basidiomycetes produce alcohol dehydrogenase, so it is feasible to produce alcohol from basidiomycetes. Agaricus blazei, Flammulina velutipes and Tricholoma matsutake have been used for mushroom fermentation to produce alcohol. To investigate whether Pholilta nameko can be used for mushroom fermentation, and find out the relationship between mannitol-1-phosphate dehydrogenase and alcohol dehydrogenase, we cloned mannitol-1-phosphate dehydrogenase gene from P. nameko, which is a zinc-containing long- chain alcohol dehydrogenase. Mpd, the gene encoding mannitol-1-phosphate dehydrogenase (MPD), has been sequenced and characterized from basiodiomycete P. nameko. The length of the coding region is 1360bp. The gene encodes a putative protein of 359 amino acids; predicted protein molecular weight is 38.6 kDa and an isoelectric point is PI = 7.34. The locations of exons and introns in the gene were deduced on the basis of interruptions in the amino acid sequence that were homologous to those in the MPD of Laccaria bicolor, the coding region was split into 6 exons and 5 introns. The protein deduced from the gene MPD showed more than 46% sequence identity to 20 fungal MPDs or alcohol dehydrogenases documented in the Gene bank protein database, based on BLASTP analysis, and was phylogenetically close to the MPDs of L. bicolor and Coprinopsis cinerea. This protein shared the same conserved domain with the alcohol dehydrogenase.

Early diagnoses of pregnancy for animal such as swine and bovine is extremely important to increase income of a farmhouse and for the management of farm. For the development of immunoasaay system of pregnancy in swine, we report a competitive heterologous enzyme linked immunosorbent assay (ELISA) for the direct measurement of oestrone sulfate (E1S) in diluted urine using anti-E1G (glucuronide) monoclonal antibody which cross react with E1S. The principle of assay was based on the typical solid-phase competitive ELISA methods using E1G-HRP (horseradish peroxidase) as a tracer and E1S for standard. The method had a reasonable sensitivity for the detection of E1S with 0.15 ng/ml as a detection limit. The intra-assay and inter-assay precisions were raging coefficient of from 8.50～9.67% and 8.50～9.87%, respectively, which were quite acceptable. In a field trial with a group 37 sows (18 non-pregnancy and 19 pregnancy sows) after day 29～30 post service, the concentration of E1S were determined to be below 30 ng/ml in all non-pregnancy group and over 48 ng/ml in pregnancy group except one sample. The method described here, heterologous ELISA for the measurement of E1S in urine is good enough for monitoring the early pregnancy test of swine.

Tissue-specific promoters are a very useful tool for manipulating gene expression in a target tissue or organ; however, their range of applications in other plant species has not been determined, to date. In this study, we identified two late pollen-specific rice promoters (ProOsLPS10 and ProOsLPS11) via meta-anatomical expression analysis. We then investigated the expression of both promoters in transgenic rice (a homologous system) and Arabidopsis (a heterologous system) using ProOsLPS10 or ProOsLPS11::GFP-GUS constructs. As predicted by microarray data, both promoters triggered strong GUS expression during the late stages of pollen development in rice, with no GUS signals detected in the examined microspores and sporophytic tissues. Interestingly, these promoters exhibited different GUS expression patterns in Arabidopsis. While in Arabidopsis, the OsLPS10 promoter conferred GUS expression at the uni- and bi-cellular macrospore stages, as well as at the shoot apical region during the seedling stage, the OsLPS11 promoter was not active in the pollen at any stage, or in the examined sporophytic tissues. Furthermore, by performing a complementation analysis using a sidecar pollen (scp) mutant that displays developmental defects at the microspore stage, we found evidence that OsLPS10, which can be an applied promoter expressed in Arabidopsis, is useful for directing gene expression in the early stages of pollen development. Our results indicate that the OsLPS10 and OsLPS11 promoters can drive the expression of target genes during the late stages of pollen development in rice, but not in Arabidopsis. Our results also emphasize the necessity of confirming the applicability of an established promoter to heterologous systems.

OsLPS is pollen specific gene that express at late stage of pollen development in rice. Based on microarray database, promoter region of two genes Os03g0106900 and Os03g0106500 were identified. The sequence of 2287bp and 2468bp upstream region of these genes were amplified and designated as OsLPS10 and OsLPS11. These promoters were fused with GUS-GFP reporter gene in a destination vector, pKGWFS7 and introduced into rice (Dongjin cultivar) and Arabidopsis (Col-0). The results of GUS assay showed different pattern of gene expression in pollen of rice and Arabidopsis. In Arabidopsis, the OsLPS10 gene strongly activated in young anther and not expressed in mature pollen. Pollen development analysis revealed GUS expression was detected at unicellular stage and strongest at the bicellular pollen developmental stage. No GUS signal was recorded in mature pollen. In case of OsLPS11, no GUS signal was detected in during pollen development of inflorescent. By contrast, in rice, the GUS expression pattern of OsLPS10 and OsLPS11 exhibited similar. GUS expression was first detectable in the anthers of spikelets at the bicellular stage and intensity increased in tricellular and mature pollen. The GUS signal was not detected in the anthers in unicellular microspores in both genes, OsLPS10 and OsLPS11. The results suggested that these genes were different activity in heterologous plant system, monocot and dicot. Complementation analysis and Cis-regulatory elements will be examined to illuminate the characteristic of these genes

We investigated Arctic plants to determine if they have a specific mechanism enabling them to adapt to extreme environments because they are subject to such conditions throughout their life cycles. Among the cell defense systems of the Arctic mouse-ear chickweed Cerastium arcticum, we identified a stress-responsive dehydrin gene CaDHN that belongs to the SK5 subclass and contains conserved regions with 1 S-segment at the N-terminus and 5 K-segments from the N-terminus to the C-terminus. To investigate the molecular properties of CaDHN, yeast were transformed with CaDHN. CaDHN-expressing transgenic yeast (TG) cells recovered more rapidly from challenge with exogenous stimuli, including oxidants (hydrogen peroxide, menadione, and tert-butyl hydroperoxide), high salinity, freezing and thawing, and metal (Zn2+), than wild-type (WT) cells. TG cells were sensitive to copper, cobalt, and sodium dodecyl sulfate. In addition, the cell survival of TG cells was higher than that of WT cells when cells at the mid-log and stationary stages were exposed to increased ethanol concentrations. There was a significant difference in cultures that have an ethanol content >16%. During glucose-based batch fermentation at generally used (30℃) and low (18℃) temperatures, TG cells produced a higher alcohol concentration through improved cell survival. Specifically, the final alcohol concentrations were 13.3% and 13.2% in TG cells during fermentation at 30℃ and 18℃, respectively, whereas they were 10.2% and 9.4%, respectively, in WT cells under the same fermentation conditions. An in vitro assay revealed that purified CaDHN acted as a reactive oxygen species (ROS)-scavenger by neutralizing H2O2 and a chaperone by preventing high temperature-mediated catalase inactivation. Taken together, our results show that CaDHN expression in transgenic yeast confers tolerance to various abiotic stresses by improving redox homeostasis and enhances fermentation capacity, especially at low temperatures (18℃).

The coat protein (CP) gene of the G5H and G7H strains of Soybean mosaic virus (SMV) were cloned, sequenced and transformed into the tobacco plant (Nicotiana tabacum. cv. Havana SR1) via Agrobacterium-mediated transformation. Transformation was confirmed b