본 리뷰의 목적은 벼 종자 저장단백질 구조분석 및 발현특성분석 결과 종합화를 통하여 종자형질 개선 등의 실용화연구를 위한 기반구축을 모색하는데 있다. 최근 벼 염색체염기서열완전해독 연구 결과를 이용한 유용형질 유전자 분리 및 실용화 연구가 많이 진행되고 있다. 특히 벼 종자 저장단백질은 인류에게는 주요 영양원으로 사용되어지며 종자 발아시에는 식물체 성장을 위한 질소원으로 사용되어진다. 벼 종자 저장단백질의 분류는 용매에서의 용해도에 따라 약산성 및 알카리 용해성의 glutelin, 알코올 용해성의 prolamin, 염 용해성의 globulin으로 나눈다. 벼 염색체 상에는 11개의 glutelin 유전자와 33개의 prolamin 유전자가 존재하며 prolamin 유전자의 경우 5번 염색체 15 Mb 부위에 15개의 유전자가 위치하였다. 이와 같이 종자저장단백질 유전자들이 동일 염색체 부위에 위치하고 있는 것은 진화학적으로 동일 염색체에서 유래하였거나 유사한 유전자발현 조절영역을 가지고 있음을 의미한다. Globulin 유전자는 5번 염색체에 단일 유전자로 존재하였다. 마이크로어레이를 이용한 종자저장 단백질 관련 유전자의 조직 특이 발현 양상을 분석한 결과 glutelin과 대다수의 prolamin 합성 유전자는 종자배유에서만 발현을 하였으며 소수의 prolamin과 globulin 합성 유전자는 종자배유와 발아종자에서도 발현을 나타내었다. 종자 저장단백질의 프로모터부위를 분리한 후 종자에서의 발현 양상을 분석한 결과 glutelin type C1 프로모터가 종자의 전체 부위에서 발현을 나타내었으며 glutelin type B5와 α-globulin 프로모터가 많은 양의 발현을 나타내었다. 본 리뷰를 통하여 벼 종자 저장단백질의 구조및 발현특성 연구 진행사항을 살펴보았다. 이러한 연구 동향분석은 종자형질 개선 및 물질생산 등의 실용화 연구를 수행하는 연구자들에게 최근의 연구 현황을 제공할 수 있을 것으로 생각된다.

Ceriporiopsis subvermispora is a unique white rot fungus degrading plant cell wall lignin without severe loss of cellulose. Recombinant plasmids containing homologous gene expression signals fused to the coding sequence of Escherichia coli hph which encodes for hygromycin phosphotransferase were introduced in protoplasts of wild type C. subvermispora using PEG/CaCl2 protocol. A number of hygromycin B resistant strains were isolated on a screening plate containing 100㎍/ml of hygromycin B: whereas, no colonies were observed when protoplasts were treated with no DNA as a negative control. It was demonstrated that most of the isolates lost their drug resistance during successive cultivations in the presence of the same concentration of hygromycin B, but some of the isolates showed stable drug resistance after five times repeated screening. They did not lose the drug resistance even after the cultivation in the absence of hygromycin B and incorporation of the hph sequence was confirmed by specific amplification of the target sequence in PCR experiments and Southern hybridization analysis. The stable transformation system will make it possible to do molecular genetic analysis, as well as breeding of genetically modified strains, in C. subvermispora. Moreover, it was demonstrated that recombinant constructs with truncated promoter showed reduced number of the drug resistant isolates on the first screening plate, in response with the length of the remaining promoter sequence. These findings indicated that unstable drug resistance observed in these isolates should originate from transient expression of the introducing marker genes, and that a promoter assay system has been developed for the first time in basidiomycete. This system is practically not affected by the positional effect of the integrated recombinant gene in the host chromosome.

Decorin (DCN) is a member of small leucine‐grich proteoglycans which are ubiquitous components of the extracellular matrix. It regulates many physiological processes, such as matrix formation, collagen fibrillogenesis, angiogenesis, cancer growth, and cardiovascular diseases. It has been shown that DCN is expressed in the uterus during pregnancy and modulates implantation and decidualization for the establishment and maintenance of pregnancy in mice and humans. Expression of DCN in the uterine endometrium during pregnancy has not been investigated in pigs. Thus, this study investigated expression of DCN in the uterine endometrium during the estrous cycle and pregnancy in pigs. Uterine endometrial tissues were from day (D) 12 and 15 of the estrous cycle and D12, D15, D30, D60, D90, and D114 of pregnancy. Northern blot and real‐gtime RT‐gPCR analyses showed that expression of DCN mRNA was detected throughout the estrous cycle and pregnancy with the highest levels during mid pregnancy. In situ hybridization analysis showed that DCN mRNA was localized to both luminal and glandular epithelia during the estrous cycle and pregnancy and also to chorionic membrane during mid pregnancy in pigs. To determine whether endometrial expression of DCN was affected by the somatic cell nuclear transfer (SCNT) procedure, DCN mRNA levels in the uterine endometrium from gilts with SCNT embryos on D30 of pregnancy were compared with those from gilts with normal embryos using real‐gtime RT‐gPCR analysis. The result showed that DCN mRNA levels in the uterine endometrium were not significantly different between gilts with normal embryos and SCNT embryos. These results suggest that DCN may play an important role for endometrial tissue remodeling during mid pregnancy, and DCN expression is not affected by the SCNT procedure at the early stage of pregnancy in pigs.

Matrix metalloproteinases (MMP) play important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, oocytes development and ovulation. In an attempt to investigate the effect of MMP activation in development cumulus-oocytes complexes, we examined the localization and expression of MMP, and monitored MMP expression profile. Cumulus-oocytes complexes were collected and matured in vitro for 24 hr, 36 hr and 48 hr. A mRNA expression of MMP-2, MMP-9, TIMP-2 and TIMP-3 was detected in all culture medium regardless of CC, OC and COCs. Activity of MMP-2 in the OC progressively was increased from 24 hr to 48 hr. But MMP-9 was not detected in all culture medium. The localization of MMP-2 was also measured by immunohistochemistry analysis. The MMP-2 and TIMP-2 was detected in cumulus cell and oocyte zone pellucida. Expression of MMP-2 protein in the COCs was progressively increased from 24 hr to 48 hr. However, MMP-9 protein was progressively decreased from 24 hr to 48 hr. And TIMP-2 protein was most highly expressed in the COCs 36 hr. Expression of TIMP-3 protein in the COCs was progressively increased from 24 hr to 48 hr. In conclusion, these results suggest that MMP-2 plays a role in maintaining normal maturation and development by controlling the ECM inhibitor concentration on cumulus cell and oocytes.

Glandular odontogenic cyst (GOC) is a rare odontogenic cyst, which shows cystic structures lined by stratified squamous epithelium with various thickness. Glandular duct-like spaces lined by eosinophilic cuboidal or columnar cells constitute the epithelial surface. The purpose of this study was to present 7 cases of GOC retrieved from the files of the Department of Oral Pathology, Seoul National University Dental Hospital and to investigate the immunohistochemical expression of cytokeratins (CKs) in the epithelial components. A total of 7 GOCs were reviewed clinically and radiographically and immunostainning for CK 5, 7, 14, 18 and CK-pan were performed. There were five males and two females aged from 36 to 53 years (mean 45 years). Maxilla was more affected than mandibles (6:1). Radiographically, all cases showed multilocular radiolucencies with well-defined borders. Histologically, lining epithelium of GOC was composed of nonkeratinized stratified epithelial cells with focal plaque-like or whirl pool-like thickenings. Surface epithelial layer contained eosinophilic cuboidal cells or mucous cells. Mucin pools of microcystic areas was also detected in the epithelium. Immunohistochemical study demonstrated that epithelium of GOCs was positively reactive for CK 5 7, 14 and CK-pan with a slight variation in their patterns and there was no reaction for CK 18.

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.

Aujeszky’s disease (AD), also called pseudorabies, is an infectious viral disease, caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. AD affects many countries and regions in the world, causing important economic losses, mainly due to international trade restrictions. In this study, to determine the characteristics of the Aujeszky’s disease virus (ADV), NYJ strain, which was isolated from the serum of an infected pig in 1987, we investigated the nucleotide sequence and expression of the glycoproteins gB, gC, and gD using the bBpGOZA system. We found that the glycoproteins gB, gC, and gD of NYJ consisted of 2751 bp, 1443 bp, and 1203 bp, respectively. Comparison of the NYJ with the other strains revealed nucleotide sequence identity ranging from 91.tito 99.0%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The NYJ strain was formed a distinct branch with high bootstrap support. The expression of glycoprotein gD in insect cells was characterized by SDS-PAGE and Western blotting with an anti-ADV polyclonal antibody. Glycoprotein gD of approximately 45 kDa was detected. The results of this study have implications for both the taxonomy of ADV and vaccine development.

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.

Lysophosphatidic acid (LPA), a simple phospholipid-derived mediator implicated in diverse biological actions, acts through the specific G-protein coupled receptors, LPA receptor (LPAR) 1～5. Our previous study showed that LPAR3 is expressed in the uterine endometrium in a cell type- and stage-specific manner and LPA via LPAR3 increases PTGS2 expression in the uterine endometrium during the period of implantation. Although LPAR3 is considered to be predominant LPA receptor in the uterine endometrium, other LPA receptors might play a role to mediate LPA functions in the uterine endometrium during pregnancy. Among LPARs, we investigated expression of LPAR1 during the estrous cycle and pregnancy in this study. Uterine endometrial tissue samples were collected from day (D) 12 and D15 of the estrous cycle and from D12, D15, D30, D60, D90 and D114 of pregnancy. Northern blot analysis determined that LPAR1 mRNA was constitutively expressed in the uterine endometrial tissues during the estrous cycle and pregnancy of all stages. Analysis by immunoblotting revealed that LPAR1 proteins were present in the porcine uterine endometrium during the estrous cycle and pregnancy. Immunohistochemical experiments demonstrated that LPAR1 protein was localized to endometrial epithelium and stromal cell, specifically to nuclei of these cell types. Results in this study show that LPAR1 is constitutively expressed in the uterine endometrium during the estrous cycle and pregnancy. These results suggest that LPA via LPAR1 may play a role in the uterine endometrial function throughout pregnancy in pigs.

In the present study, we have used an annealing-control-primer (ACP)-based differentially display RT-PCR method to identify salt-stress-induced differentially expressed genes (DEGs) in barley leaves. Using 120 ACPs, a total of 11 up-regulated genes were identified and sequenced. Temporal expression patterns of some up-regulated DEGs in response to salt stress were further analyzed by Northern blot analysis. The possible roles of these identified genes are discussed within the context of their putative role in response to salt stress. Thus, the identification of some novel genes-such as SnRK1-type protein kinase; 17 kDa, class I, small heat shock protein; and RNase S-like protein precursor genes-may offer a new avenue for better understanding the salt stress response in plants, knowledge which might be helpful for developing future strategies.

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.