This study aimed at investigating the gene expression profile in basal ganglia of hexa-valence chromium exposed rat based on cDNA array analysis. For cDNA array, Sprague-Dawley male rats (300 ± 25 g) were administrated with 15 mg/kg B.W/day of potassium dichromate by gavage (0.3 ml) dissolved in saline for 10 days (n=5). For dose-related gene expression analysis, rats were administrated with 0, 1, 5 mg/kg B.W/day of potassium dichromate for 10 days. Control rats were administrated with equal volume of saline (n=5). For cDNA array analysis, RNA samples were extracted from brain tissue and reverse-transcribed in the presence of [α³²P]-dATP. Membrane sets of the Atlas array II and Toxicology array kits were hybridized with cDNA probe sets. RT-PCR and Northern blot hybridization methods were employed for validation and assessment of the dose-related gene expression profile, respectively. Among the 2352 cDNAs, 43 genes showed significant (>two-fold) changes in expression. 22 genes were up-regulated and 21 genes were down-regulated in the 15 mg/kg B.W/day hexa-valence chromium treated group than control. According to the Northern blot hybridization analysis, heat shock protein 47, neurodegeneration associated protein 1 and pituitary specific growth factor 1a genes were up-regulated, but Gamma-aminobutyl-acid a1 subunit, neuroligin2, brain calcium-transporting plasma membrane type ATPase genes were down-regulated even in the low-dose of hexa-valence chromium exposed group (1 mg/kg B.W/day) than control. Genes that detected in this study may be closely related to the hexa-valence chromium-induced neurotoxicity in the rat basal ganglia and addition study of these genes can give some more useful information about the neuro-toxic mechanism by hexa-valence chromium.

The miniature pig is considered to be a better organ donor breed for xenotransplantation than other pig breeds because the size of the organs of the miniature pig is similar to that of humans. In this study, we aimed at identifying differentially expressed genes in the miniature pig ovary during pregnancy. For this, we used the miniature pig ovary model, annealing control primer‐based reverse transcription polymerase chain reaction (PCR), quantitative real‐time PCR (qRT‐PCR), and northern blotting analysis. We identified 13 genes showing differential expression on the based of pregnancy status and validated 8 genes using qRT‐PCR. We also sequenced the full‐length cDNA of ephrin receptor A4 (EphA4), which had a significant difference in expression level, and validated it by northern blotting. These genes may provide a better understanding of the cellular and molecular mechanisms during pregnancy in miniature pig ovary.

To determine differential gene expression profiles in the venom gland and sac (gland/sac) of a solitary hunting wasp species, Eumenes pomiformis Fabricius (1781), a subtractive cDNA library was constructed by suppression subtractive hybridization. A total of 541 expressed sequence tags (EST) were clustered and assembled into 102 contigs. In total, 37 genes were found from the library by BLASTx search and manual analysis. A eumenitin-like venom peptide, EpVP1, occupied ca. 26% of the library. A novel venom peptide, EpDTX, shared sequence similarity with trypsin inhibitors and dendrotoxin-like venom peptides known as K+ channel blockers, implying it could be responsible for the paralysis of prey. As well as phospholipase A2 and hyaluronidase known to be main components of wasp venoms, several contigs encoding enzymes, including metalloendopeptidases and a decarboxylase likely involved in the processing and activation of venomous proteins, peptides, and neurotransmitters, were also isolated from the library. The presence of a gene encoding insulin-like growth factor binding protein suggests that solitary hunting wasps might control the prey to stay in larval stage by their venom. The abundance of these venom components in the venom gland/sac and alimentary canal was confirmed by quantitative real-time PCR.

Many biological systems are regulated by an intricate set of feedback loops that oscillate with a circadian rhythm of roughly 24 h. This circadian clock mediates an increase in body temperature, heart rate, blood pressure, and cortisol secretion early in the day. Recent studies have shown changes in the amplitude of the circadian clock in the hearts and livers of streptozotocin (STZ)-treated rats. It is therefore important to examine the relationships between circadian clock genes and growth factors and their effects on diabetic phenomena in animal models as well as in human patients. In this study, we sought to determine whether diurnal variation in organ development and the regulation of metabolism, including growth and development during the juvenile period in rats, exists as a mechanism for anticipating and responding to the environment. Also, we examined the relationship between changes in growth factor expression in the liver and clock-controlled protein synthesis and turnover, which are important in cellular growth. Specifically, we assessed the expression patterns of several clock genes, including Per1, Per2, Clock, Bmal1, Cry1 and Cry2 and growth factors such as insulin-like growth factor (IGF)-1 and -2 and transforming growth factor (TGF)-β1 in rats with STZ-induced diabetes. Growth factor and clock gene expression in the liver at 1 week post-induction was clearly increased compared to the level in control rats. In contrast, the expression patterns of the genes were similar to those observed after 5 weeks in the STZ-treated rats. The increase in gene expression is likely a compensatory change in response to the obstruction of insulin function during the initial phase of induction. However, as the period of induction was extended, the expression of the compensatory genes decreased to the control level. This is likely the result of decreased insulin secretion due to the destruction of beta cells in the pancreas by STZ.

Perennial ryegrass (Lolium perenne L.) is one of the most important grass species in the world's temperate zones. It is used as high-quality forage in pastures and for recreational use as turf in golf courses, lawns and parks. Genetic improvement of perennial ryegrass is difficult due to its self-incompatibility. Consequently, progress by conventional breeding can be slow. Genetic transformation is an alternative that permits direct introduction of useful genes into a plant's genome and is becoming a powerful tool to complement conventional breeding. To improve environmental stress tolerance and quality of perennial ryegrass by introducing better and useful genes into the genome, we have developed a rapid and efficient transformation system using Agrobacterium-mediated gene transfer system.

To understand molecular and cellular mechanisms of many gene products in the female reproductive organs including the ovary and uterine endometrium as well as during embryo development, researchers have developed and utilized many effective methodologies to analyze gene expression in cells, tissues and animals over the last several decades. For example, blotting techniques have helped to understand molecular functions at DNA, RNA and protein levels, and the reverse transcription-polymerase chain reaction (RT-PCR) method has been widely used in gene expression analysis. However, some conventional methods are not sufficient to understand regulation and function of genes expressed in very complex patterns in many organs. Thus, it is required to adopt more high-throughput and reliable techniques. Here, we describe several techniques used widely recently to analyze gene expression, including annealing control based-PCR, differential display-PCR, expressed sequence tag, suppression subtractive hybridization and microarray techniques. Use of these techniques will help to analyze expression pattern of many genes from small scale to large scale and to compare expression patterns of genes in one sample to another. In this review, we described principles of these methodologies and summarized examples of comparative analysis of gene expression in female reproductive organs with help of those methodologies.

A full genomic DNA microarray technique was employed to investigate the effects of Dongchunghacho on aortal and hepatic gene expression in apolipoprotein E knockout mice fed a high-fat/high-cholesterol diet. Male 8- week - old ApoE-/- mice were randomly divided into two groups, control(high cholesterol group; HC) and supplementation of Dongchunghacho (SD). All of the mice were fed a high-fet/high cholesterol diet with or without Dongchunghacho supplemented by 1% for 6 weeks. At first, lipid profile of the Dongchunghacho was measured by biochemical analysis. No differences were observed in serum triglyceride and total cholesterol levels between the two groups. Antigenotoxic effect of the Dongchunghacho was measured by the single cell gel electrophoresis assay (Comet assay) and quantified as % fluorescence in tail. Dongchunghacho supplementation decreased significantly leukocytic DNA damage and also there was a tendency of reduction in hepatic DNA damage in Dongchunghacho group compared with the control group. In up regulated genes in liver and aorta of the mice, genes with 0 to 2- fold difference in expression level between the two group (HD and SD) was very much more in liver than in aorta, on the contrary, those with 2-fold to 16-flod difference increased greatly rather in aorta than in liver. Also, almost the same results were observed in down regulated genes in liver and aorta between the two groups. These results suggested that supplementation of Dongchunghacho might be helpful in preventing leukocytic DNA damage induced by high fat diet, and has a more crucial roles in aortal gene expression.

Protease from various sources have been studied biotecnologically. For biotechnological applications, one highly preferred enzyme is protease. There have been no reports of cloned genes encoding digestive proteases in the Laccotrephes japonenis, Ranatra unicolor, Muljarus japonicus. These insects are considered to be a predator of aquatic insects. RT-PCR was used to amplify cDNA fragments for digestive proteases from total RNA the hole body of the insects. The flanking sequences of the 5'- and 3'- end of the these genes were characterized by RACE-PCR. Sequence analysis showed that these genes contained complete ORF. The deduced amino acid sequences of these protease showed 62% identity to the serine protease of Creontiades dilutus, 58% to Lygus loneolaris trypsin-like serine proteinase, 54% to Triatonatoma infestans salivary trypsin. To generate Laccotrephes japonensis serine protease, the DNA fragment coding for serine protease is cloning into suttle vector pBACⅠ, named pBAC1-JG and infected to Spodoptera frugiperda (sf9) insect cell. The cDNA encoding JG was expressed as a 32-kDa polypeptide in baculovirus infected insect cells and the recombinant protein showed activity in the protease enzyme assay using gelatin as a substrate.

The use of electron beam irradiation has been emerged as one of alternative ways of the restriction of methyl bromide usage for the disinfestation of stored-products and quarantine pests. Here we demonstrated effects of electron beam irradiation on development and gene expression of P. interpunctella, which is a serious pest of various stored-products. P. interpunctella at various developmental stages were irradiated at 0, 0.25, 0.5, 0.75 and 1 kGy. Eggs failed to hatch at all treated doses of electron beam. Fifth instar larvae pupated only 6.7% by 0.25 kGy irradiation but failed pupation at above doses. Interestingly, survived larvae by low-dose irradiation did not pupated until 40 days. Pupae eclosed to adults only 12.1% by 0.25 kGy irradiation but failed at above doses. In addition, 5-day-old pupae eclosed 94.4, 91.6, 100 and 49.9% at 0.25, 0.5, 0.75 and 1.0 kGy, respectively. However, most of those emerged adults were malformed, especially in wings, and showed very low oviposition rate. We demonstrated whether the electron beam irradiation induces gene expression. Upregulated genes at any doses were hsp70, which is a stress-responsive protein, at fifth instar larval stage and hemolin, which is an immune-responsive protein, at pupal stage. Some genes of pupae, such as β-1,3 glucan recognition protein, hsp70 and acp25 (small hsp) were upregulated only at high doses. However, other genes, such as prophenoloxidase, ultraspiracle , ecdysone receptor and heat shock proteins (hsp90, hsc70) were downregulated by irradiation of electron beam

A glucose-regulated protein 78 (grp78) gene, which is belongs to a heat shock cognate 70 (hsc70) subfamily, was cloned from Indian meal moth, Plodia interpunctella. Its full length cDNA was 2679 bp and contains a 1980 bp open reading frame. The translated amino acid sequence consists of 660 residues with a calculated molecular mass of 72,975 Da and an isoelectronic point (pI) of 5.27. It contains several highly conserved functional motifs of the Hsp70 family and, particularly, C-terminal motif of KDEL that is characteristic for endoplasmic reticulum (ER) hsc70. Its deduced amino acid sequence shows a high identity (83-94%) with Hsc70s of other insects and grouped with Hsc70-3 among 5 Hsc70 members of Drosophila melanogaster. During development the grp78 transcript level was high in egg, feeding larval and adult stages but low in molting and wandering larval and pupal stages. Particularly its level was higher in the gut than integument and fat body of fifth instars. Furthermore its level was greatly decreased when fifth instar larvae were starved for 48 hrs but recovered at 3-6 hrs after re-fed diet. Our data suggests that grp78 is a member of hsc70 gene that belongs to ER and may have a role for energy metabolism at cellular level.