The small brown planthopper (SBPH), Laodelphax striatellus, is one of the most serious pest insects of rice plants because it can transmit the rice stripe virus (RSV) which often causes significant reduction of yield in the field. Buprofezine is an effective insect growth regular (IGR) pesticide to control planthoppers, however, since the use of buprofezine for more than a decade, it has caused a certain resistance of SBPH. To survey the responses of SBPH to buprofezine, we exposed 4th instar SBPH to 200 ppm buprofezine by dipping method, and extracted total RNA for RNA-seq by Illumina platform. The quality filtered raw reads of cDNA obtained from experimental and control SBPH were subjected to Bowtie2 followed by eXpress computer program to compare the differential gene expression which will be important information for pest control methods using RNAi.

Differential gene regulation is crucial for development. Indianmeal moth is a global pest of stored and processed food products. Here, we determined the whole developmental expression patterns of 13 genes (shsp, hsp70, grp78, and hsp90), ecdysone receptor (EcR), ultraspiracle (USP), hemolin, β-1,3-glucan recognition protein (βgrp), prophenoloxidase (ProPO), superoxide dismutase (SOD) and thioredoxin peroxidase (Tpx), and two hexamerin storage protein genes (SP1 and SP2) related to growth, stress, metabolism, and host defense. We also studies effects of Bracon hebetor envenomation on these gene expressions to explore their role in host regulation. We found unexpected transcriptional peaks especially hsps which were high in egg and adult stages. This study provides comprehensive understanding about development and parasitoid regulation at molecular level.

Clematis patens는 미나리아재비과 으아리속에 속하는 다년 생 숙근초로 꽃잎화한 꽃받침을 가지고 있는 점이 특징이며 흰색, 분홍색, 자주색, 보라색, 등 다양한 화색이 존재한다. 본 연구에서는 C. patens ‘The President’에서 flavonoid 생합성 경로에 관여하는 유전자 중 delphinidin 계열 anthocyanin 색 소 합성을 유도하는 F3'5'H(ClF3'5'H)를 분리 동정하였으며, 화색 및 발달 단계별 ClF3'5'H 유전자의 발현 패턴을 분석하 였다. 또한 클레마티스 화색별 anthocyanin의 함량을 알아보 았다. 그 결과, 기존 보고된 Aconitum carmichaelii의 F3'5'H 유전자의 아미노산 서열과 79% 일치하여 높은 상동성을 갖는 것을 확인하였으며, ClF3'5'H 유전자의 발현 패턴은 흰색을 띠 는 클레마티스에서는 ClF3'5'H 유전자의 발현량이 많지는 않 았으나 다른 발달 단계에 비해 완전 개화한 단계에서 발현이 강했다. 분홍색을 띠는 클레마티스에서는 모든 발달 단계에서 ClF3'5'H 유전자의 발현이 검출되지 않았으며, 자주색과 보라 색을 띠는 클레마티스에서는 개화 직전 및 완전 개화 단계에 서 발현하였다. 이 결과로부터 클레마티스의 다양한 화색과 클 레마티스의 F3'5'H 유전자 발현과의 상관관계가 시사되었다.

Transcription factor called activating enhancer binding protein 2C (AP2-gamma) is found in a variety of species and expressed from oocyte stage onwards, particularly restricted to the trophectoderm. Recent studies demonstrated that ablation of Tfap2c led to failure of tight junction biogenesis, particularly the knock-down embryos of Tfap2c did not form cavity from morula to blastocyst in mouse and pig. We speculated that the Tfa2pc may also be involved in desmosome biogenesis because blastocoel formation is coincident with the establishment of desmosome. To determine this, we depleted Tfap2c injecting siRNA into one-cell zygote and analysed the expression levels of genes that are required for desmosome complex such as PkP2, Pkp3, Dsc2, and Dsg2. We found only Pkp3 was up-regulated in the knockdowned morula embryos. Interestingly, upstream region of Pkp3 had putative Tfap2c binding sites. In conclusion, our results suggest that Tfap2c is not a crucial factor but somehow it might be involved in desmosome biogenesis directly or indirectly via Pkp3.

Chitin synthase (CHS) is an important enzymatic component, which is required for chitin formation in the cuticles and cuticular linings of other tissues in insects. CHSs have been divided into two classes, class A and B, based on their amino acid sequence similarities and functions. Class A CHS (CHS-A) is specifically expressed in the epidermis and related ectodermal cells such as tracheal cells, while expression of class B CHS (CHS-B) is expressed in gut epithelial cells that produce peritrophic matrices. In this study, we cloned the CHS-A gene from the beet armyworm, Spodoptera exigua. The SeCHS-A mRNA was expressed in all developmental stages and specifically in the epidermis and tracheae tissue by RT-PCR analysis. Expression of SeCHS-A gene was suppressed by feeding double-stranded RNA (dsRNASeCHS-A, 150 ng/larva) in the fifth instar of S. exigua. The suppression of SeCHS-A gene expression significantly induced mortality on pupal stage. Also, larvae fed with dsRNASeCHS-A significantly enhanced pathogenicity of an entomopathogenic fungus, Beauveria bassiana ANU1. These results suggest that the SeCHS-A gene plays an important role in development of S. exigua and dsRNA, which is a specific to SeCHS-A gene, may be applied to effective pest control with B. bassiana.

Pheromone biosynthesis-activating neuropeptide (PBAN), produced in subesophageal ganglion, is known to stimulate pheromone biosynthesis in Plutella xylostella. The pheromone production is more active in the scotophase than in the photophase, which suggests that there may be changes of gene expression in the pheromone glands. To analyze gene expression involving in pheromone biosynthesis for 24 hrs, we performed transcriptomes of pheromone glands which were isolated at every 4 h. Eleven pheromone biosynthesis-related genes, acetyl-CoA carboxylase (ACC), fatty acid synthase (FAS), acyl-CoA dehydrogenase, enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, Δ9 desaturase, Δ11 desaturase, fatty acid reductase (FAR), alcohol oxidase, aldehyde oxidase, and aldehyde reductase were identified. Among these genes, ACC, FAS and chain shortening enzymes involving in early stage of pheromone biosynthesis exhibited their highest expression between AM9 and PM5. Desaturases, Δ9 and Δ11, showed the peak of expression at PM1 and AM5 or PM5, respectively. Interestingly, FAR expression was the highest at AM1, active reproductive period. These results suggest that genes involving in pheromone biosynthesis can be sequentially regulated for their biological roles.

Human tissue-type plasminogen activator (t-PA) is responsible for fibrin-specific plasminogen activation and plays a key role in fibrinolysis thereby aiding breakdown of blood clots in the vasculature. In the present study, in order to develop a system for production of recombinant st-PA and t- PAHis6 proteins in transgenic rice seeds, a DNA fragment encoding t-PA gene was selected and cloned to a plant binary vector (pMJ21) harboring a rice GluB1 promoter, an N-terminal signal peptide of the rice glutelin B1 protein and a Pin II terminator. The constructed plasmid was transformed into Agrobacterium tumefaciens LBA4404 (pSB1) to facilitate introduction into rice callus. The insertion of the st-PA and t-PAHis6 genes into the genome of transgenic rice seeds and their transcripts were confirmed using PCR, and Southern blot as well as RT-PCR, respectively. The highest level of recombinant st-PA expression as determined by enzyme-linked immunosorbent assay (ELISA) was found to be 2,916 ng/total soluble protein (mg) in transgenic rice seeds. The amount of recombinant proteins expressed in transgenic plants was estimated to range from 634 ~ 2,916 ng/TSP mg (st-PA) and 925 ~ 2,640 ng/TSP mg(t- PAHis6), respectively. Immuno-blot analysis of transgenic rice seeds revealed single bands of approximately 68-kDa representing recombinant st-PA and t-PAHis6 proteins. These results demonstrate the expression and in vivo activity of recombinant st-PA and t-PAHis6 in transgenic rice seeds. This study is a promising endeavor for production of recombinant pharmaceutical proteins using rice seed system.

Environmental changes exert harmful effects on organisms inhabiting coastal regions. These changes are also associated with reduced production in aquaculture farms. In this study, we investigated internal and external responses of two Bivalvia species (Crassostrea gigas and Mytilus galloprovincialis) in Gamak Bay under stressful environmental conditions in aquaculture farms. We investigated external responses such as weight, size, and environment exposure time, and analyzed the expression of the HSP70 gene. C. gigas HSP70 gene expression level was significantly high in the C3 aquaculture farm site, but the weight and size of C. gigas were high in the C2 aquaculture farm site. The response of C. gigas HSP70 mRNA was associated with the environmental exposure time in each aquaculture farm. Expression of M. galloprovincialis HSP70 gene was found to be significantly higher in the M2 aquaculture farm site than in the M1 site, whereas the weight of M. galloprovincialis was observed to be higher in the M1 site. The size and environmental exposure time of M. galloprovincialis were similar between M1 and M2 sites. In addition, HSP70 sequences of C. gigas and M. galloprovincialis showed high similarity with that of another marine species. According to our results, there were differences in internal responses following environmental stress in aquaculture farms, with respect to HSP70 gene expression. The results suggest that the HSP70 gene is a useful molecular indicator for monitoring stress responses in Bivalvia species in the field.

밀싹 추출물의 생리활성 및 화장품 소재로서의 가능성 여부를 규명하고자 하였다. 본 연구는 세포실험을 통해 밀싹 추출물의 피부 세포에 대한 독성을 확인하고, 피부 세포 미백 활성 및 노화에 대 한 연구를 수행하여 기능성 화장품 소재로서의 가능성을 알아보고자 하였다. 본 연구 결과 밀싹 추출물 이 HDF, B16F10 세포에 대한 독성이 적은 것으로 확인되었다. 멜라닌 생합성 억제에 대한 효과는 약 한 것으로 확인되었으나, 자외선에 유도되는 MMP-1의 발현을 저해함으로써 피부의 광노화를 억제하는 효과를 통해 광노화에 의한 피부 주름과 자연 노화에 의한 피부 주름을 예방하는데 유의한 효과를 가질 수 있음을 확인하였다. 따라서 본 연구는 밀싹 추출물의 기능성 화장품 소재로 사용 시 피부 노화 예방 관점에서의 기능성 화장품 소재로 매우 유용하게 활용될 수 있을 것으로 사료된다.

Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat β-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.

Obesity is a risk factor for various diseases, including cardiovascular disease, diabetes, renal disease, hypertension, cancer, and neural disease. Adipose tissue in animals is important for the mobilization of lipids, milk production, deposition of fat in different depots, and muscle and meat production. Understanding the genetic and physiological causes of metabolic disease is a priority in biomedical genome research. In this study, we examined several variables in mice fed a high-fat diet, including serum composition, body weight, total calorie intake, and differentially expressed genes. Body weight and blood glucose levels were not significantly different between animals fed high-fat and normal diets. However, high-fat diet groups showed reduced calorie and food intakes. Levels of sodium, ionized calcium, glucose, hematocrit, hemoglobin, pH, PCO2, PO2, TCO2+, HCO3+, base excess, and SO2 in the blood were not significantly different between mice fed high-fat and normal diets. Serum potassium concentration, however, was lower in mice a high-fat diet. Differentially expressed genes were also compared between the two groups. The purpose of this study was to discover new genes as a result of annealing control primer (ACP) PCR using 20 random primers. Five down regulated genes were identified and three of others were up-regulated by high-fat diet. Known genes were excluded from this result. In addition, the relationships among candidate genes and high-fat diet should be investigated according to potassium concentration in the blood. In conclusion, mice fed normal and high-fat diets showed no significant difference in body weight, whereas high-fat diet led to changesin blood composition and differential expression of several genes. These findings may provide a better understanding of the mechanisms underlying the association between obesity and metabolic diseases.

복숭아 네 품종 ‘오도로끼’, ‘가납암백도’, ‘진미’, ‘장호원황 도’ 신초의 저온순화 및 탈순화 동안 시기별 내한성 변화는 전해질 누출률을 분석하여 나타냈다. 또한 내한성 결정 요인 을 분석하고자 SDS-PAGE를 이용하여 dehydrin 함량 변화를 확인하였으며, 그와 관련된 유전자 발현 분석은 quantitative real-time RT-PCR을 이용하여 수행하였다. 네 품종의 내한성 은 2012년 12월까지 꾸준히 증가하였으며 그 후 2013년 4월 까지 감소하였다. PpDhn1 유전자가 인코딩하는 60kDa의 dehydrin 단백질은 탈순화기(2013년 3-4월)에 비하여 저온순화 기(2012년11월-2013년 1월) 동안 높은 축적이 확인되었다. PpDhn1 유전자와 PpDhn3 유전자 발현양상은 복숭아 네 품 종에서 내한성 변화와 평행하게 나타난 반면, PpDhn2 유전자 는 뚜렷한 시기별 패턴을 나타내지 않았다.

Insect Growth Regulators (IGRs) are insecticides that disrupt the normal development of target insects by inducing symptoms such as premature molting or supernumerary larval stages. Juvenile hormone systems become the targets of two types of IGRs: the Juvenile Hormone Agonists (JHAs) and Juvenile Hormone Antagonists (JHANs). Pyriproxyfen is one of the chemical compounds widely used as JHA to control many kinds of insects while Kanakugiol is a plant-extracted compound which acts as JHAN. The small brown planthopper, Laodelphax striatellus, is one of the most serious pest insects of rice plants because it can transmit the rice stripe virus which often causes significant reduction of yield in the field. In order to analyze the differential gene expressions of L. striatellus upon JHA and JHAN treatment by using next generation sequencing technique, we sprayed Pyriproxyfen and Kanakugiol on 4th instar nymphs of L. striatellus respectively, and extracted total RNA for RNA-seq. The quality-filtered Illumina sequence reads of the control, JHA, and JHAN treated samples were mapped to the reference gene sequences by using the Bowtie2 software. Then the results of mapping by Bowtie2 were analyzed by eXpress software to quantity the differential gene expression.