The nature of molecular mechanisms governing embryonic cell block is largely unknown, but recent reports have demonstrated that proper execution of programmed cell death is crucial for this process. The main objective of this study is to determine effects of programmed cell death on porcine oocytes development in vitro after parthenogenesis. Among the blastocysts matured in 3MA, MAP1LC3A and ATG5 RNA gene expression level increased in the order of Cyst < 3MA < RP. However, Casp-3 and TNF-r RNA gene expression level decreased in the order of RP < 3MA < Cyst. Expression of mTOR within the RP-cultured blastocyst was the most highly to the inner cell mass, while 3MA-cultured blastocyst showed very lowest expression in inner cell mass. The expression of mTOR showed a pattern opposite to that of MAP1LC3A. That is, its expression was the lowest in Cyst group. When the enzymatic activity of MMP-2 and MMP-9 was assessed in culture, the level of active MMP-9 was higher expression in the medium of each RP treatment group, with the level of another treatment group being relatively higher. Analyses of TIMP-2 and TIMP-3 revealed that their expression was higher in groups that did not receive RP treatment. More specifically, the level of TIMP-2 was not affected by Cyst treatment, while the level of TIMP-3 was higher in 3MA and RP treatment group. There was highly cell division activation efficiency of parthenogenesis on cultured system of RP supplement IVC medium. Therefore, these results suggest that embryo development was significantly increased in conditional culture medium with active autophagy as compared to common cultured condition. Further investigation of this distinction may enable the development of innovative improvements for the production of porcine somatic cell nuclear transfer.

전자코와 GC/MS 기기를 이용하여 나도풍란의 꽃에서발현되는 주요 향기성분 및 향기발현패턴을 분석하였다.전자코와 GC/MS 의 분석을 통해 전자코에서는 약 9개의 chromatogram peak 를 얻어냈고, GC/MS 기기분석에서는 약 13개 정도의 chromatogram peak를 얻어냈다. 전자코 peak 중에서 6가지 종류가 GC/MS chromatogrampeak와 공통적으로 나타났으며 즉 retention time 3.2초,4.2초, 5.4초, 5.8초, 6.3초, 6.9초의 peak였으며 그 peak에 해당되는 향기성분들은 각각 2-furanmethanol, linalool,citronellol, neral, nerodidol, benzoic acid, hexadecanoicacid, 1,2-benzenedicarboxylic acid로 추정되었다. 나도풍란군집에서 개체별로 향기발현량과 향기패턴을 비교분석한결과 주요성분으로 예측되는 6개 peak는 모든 개체에서발현되는 것으로 나타났지만 개체간 발현량에 차이가 있는 것으로 나타났다. 꽃의 발달단계별 향기발현정도를 분석하기 위한 실험에서 꽃의 발달 단계중 꽃봉오리 상태와 노화된 꽃에서는 향기발현량이 적었으며 꽃이 완전개화한 화서 중앙부위에 있는 꽃들에서 가장 많은 양의 향기가 발생되는 것으로 나타났다. 꽃의 기관별 향기분석에서는 나도풍란 꽃의 주요향기성분이 주로 sepal과 petal조직에서 가장 많이 발현되는 것으로 확인되었으며 column과 spur에서는 발생량이 매우 적은 것으로 나타났다. 일중 시간대별 주요향기성분의 발현량과 패턴을 분석한 결과 오전11시에 가장 높은 향기발현량을 보였으며 오후 5시 이후부터 향기발현량이 현저히 줄기 시작하여 저녁 8시 이후에는 향기성분이 발생되지 않았다. 빛 조사가 향기발현에 미치는 영향을 알아보기 위해 암처리와 광처리후 향기양과 패턴을 분석한 결과 암처리에서는 40시간 이후부터는 대체로 향기성분이 계속 줄어들었으며, 40시간연속적으로 빛을 조사한 후 20시에 전자코 분석을 한 결과 오전 시간대와 동일한 향기발현량과 발현패턴을 보여주었다. 이 결과를 통해 빛의 조사시간 및 생체리듬주기가 향기발현에 큰 영향을 줄 수 있는 요인임을 확인할수 있었다.

The aim of the present study was to identify coat color associated genes that are differentially expressed in mature Korean brindle cattle (KBC) with different coat colors and in Hanwoo cows. KBC calves, before and after coat color appearance, were included. Total cellular RNA was isolated from the tail hair cells and used for microarray. The number of expressed coat color associated genes/probes was 5813 in mature KBC and Hanwoo cows. Among the expressed coat color associated genes/probes, 167 genes were the coat color associated genes listed in the Gene card database and 125 genes were the pigment and melanocyte genes listed in the Gene ontology_bovine database. There were 23 genes/probes commonly listed in both databases and their expressions were further studied. Out of the 23 genes/probes, MLPH, PMEL, TYR and TYRP1 genes were expressed at least two fold higher (p<0.01) levels in KBC with brindle color than either Hanwoo or KBC with brown color. TYRP1 expression was 22.96 or 19.89 fold higher (p<0.01) in KBC with brindle color than either Hanwoo or KBC with brown color, respectively, which was the biggest fold difference. The hierarchical clustering analysis indicated that MLPH, PMEL, TYR and TYRP1 were the highly expressed genes in mature cattle. There were only a few genes differentially expressed after coat color appearance in KBC calves. Studies on the regulation and mechanism of gene expression of highly expressed genes would be next steps to better understand coat color determination and to improve brindle coat color appearance in KBC.

향기를 가진 미니다화성계 품종을 육성하기 위해 강하 고 향기를 가지는 P. violacea 와 붉은색 화색을 가지는 중륜계 품종인 P. George Vasquize 간 인공교잡을 통해 잡종후대를 육성하였다. 무균파종과 조직배양을 통해 약 280개의 잡종후대를 얻어냈으며 각각의 개체들이 향기의 강도와 종류가 달랐지만 대부분 모든 잡종개체들에서 향 기가 있었다. 개화기에 화색과 화형 그리고 향기의 유무 등을 고려하여 7개체를 1차 선발하였고 2차 개화시기에 꽃배열, 화수, 생육속도 등을 조사하여 최종적으로 향기 가 강하고 화색이 밝은 적색인 미니다화성 한 계통(06- SC-29) 을 선발 육성하였다. SPME-CG/MS를 통한 향 기 분석에서 51개의 성분을 검출하였고 그 중 myrcene, sabinene, ocimene, cineolebergamototene, cardinene 과 linalool 등이 성분함량이 높게 검출되어 이 물질들이 주 요 향기성분 인 것으로 추정되었다. 향기발현 관련유전 자를 분리하기 위해 유향성 꽃에서 mRNA를 분리 후 NGS 분석을 시도하여 염기서열정보를 확보하고 이미 알려 진 향기관련 유전자의 정보를 통해 호접란 향기발현관련 유 전자 정보를 가진 contig와unigene을 찾아내었다. 꽃향기 발 현에 중요한 역할을 하는 4개의 효소 linalool synthase(LIS), geranyl diphosphate synthase(GDPS), ocimene synthase(OS), farnesol kinase(FNK)를 coding하는 유전자 염기서열정보를 확보하여 다른 식물의 향기 유전자와 homology를 비교 하고, 꽃의 발달단계별, 꽃의 화기조직별 향기관련 유전자들에 대한 발현 정도를 검정하였다. 꽃의 발달 단계별 향기유전자는 유향종에서는 정도 차이는 있지만 거의 모 든 단계에서 발현이 관찰되었으나 무향종에서는 발현되 지 않거나 아주 약하게 발현되었다. 유향종의 화기조직 별 발현유무검정에서는 4개의 유전자가 꽃잎, 꽃받침, 암 술, 꽃술대에서 모두 고르게 발현되었다.

Phospholipase A2 (PLA2) catalyzes an ester hydrolysis at sn-2 position of phospholipids. Various PLA2 genes are classified into at least 15 groups. However, on the basis of physiological functions, PLA2 genes are classified into calcium dependent cellular PLA2 (cPLA2), calcium independent cellular PLA2 (iPLA2) and secretary PLA2 (sPLA2). In insects, several sPLA2 genes are known to be associated with venom or immune functions. However, no known cellular PLA2 genes are identified. This study reports an iPLA2 (SeiPLA2) encoded in Spodoptera exigua. SeiPLA2 has an open reading frame of 2448 bp encoding a sequence of 816 amino acid residues. Its predicted protein is 89.55 KDa and 6.15 pI. SeiPLA2 is expressed in egg, larva, pupa and adult stages. In larval stage, SeiPLA2 is expressed in hemocytes, fat body, epidermis, gut, malpighian tubules and salivary gland. To understand its physiological function, its RNA interference is under investigation.

This experiment was conducted to investigate the genetic effects of candidate genes on the growth of spleen and liver tissues using dietary Curcuma longa (C. longa) supplementation. Expression analyses of candidate genes regarding animal growth was performed in order to determine the factors affecting the growth related to immune components of Curucumin, Turmerone, and Zingiberene as the bile secretion Paratolyl methyl carbinol (PTMC). The animals were divided into four groups of five chicks supplied with experimental diets of C. longa at 0.25, 0.5 and 1% and controls. The 19 growth-related genes were known to cell maturation, differentiation significant expression patterns in this analysis. Expression of growth response-related genes in chicks supplemented with 1% of C. longa showed better growth performance than chicks with 0.25 and 0.5% in spleen (p<0.05). The IGF1, MSTN, POU1F1, ADCYAP1 gene were known to central roles in mediating gonadotropin function, regulating steroidogenesis and promoting oocyte growth and maturation. Sex steroids, androgen and estrogen can affect sex differentiation and also can affect muscle development. On the other hand, GHSR and FABP3 gene showed significant expression patterns in this analysis. The results would be used as basic information for the variation of growth-related genes expression on the cell growth, sex cell growth, and sex hormones according to dietary supplementation with C. longa in chickens.

Many countries have implemented genetic evaluation for fertility traits in recent years. In particular, reproductive trait is a complex trait and need to require a system-level approach for identifying candidate genes related to the trait. To find the candidate gene associated with reproductive trait, we applied a weighted gene co-expression network ana-lysis from expression value of bovine genes. We identified three co-expressed modules associated with reproductive trait from bovine microarray data. Hub genes (ZP4, FHL2 and EGR4) were determined in each module; they were topologically centered with statistically significant value in the gene co-expression network. We were able to find the highly co-expressed gene pairs with a correlation coefficient. Finally, the crucial functions of co-expressed modules were reported from functional enrichment analysis. We suggest that the network-based approach in livestock may an important method for analyzing the complex effects of candidate genes associated with economic traits like repro-duction.

CD63, a member of tetraspanin membrane protein family, plays pivotal role in cell growth, motility, signal transduction, host-pathogen interactions and cancer. In this work, the cDNA encoding CD63 homologue (TmCD63) was cloned from larvae of coleopteran beetle, Tenebrio molitor. The cDNA is comprised of an open reading frame of 705 bp, encoding putative protein of 235 amino acid residues. In silico analysis shows that the protein has four putative transmembrane domains and one large extracellular loop. The characteristic ‘Cys-Cys-Gly’ motif and ‘Cys188’ residues are highly conserved in the large extracellular loop. Phylogenetic analysis of TmCD63 revealed that they belong to the insect cluster with 50-56% identity. Analysis of spatial expression patterns demonstrated that TmCD63 mRNA is mainly expressed in gut and Malphigian tubules of larvae and the testis of the adult. Developmental expression patterns of CD63 mRNA showed that TmCD63 transcripts are detected in late larval, pupal and adult stages. Interestingly, TmCD63 transcript was upregulated the maximum 4.5 fold in response to DAP-type peptidoglycan during the first 6 h, although other immune elicitors also made significant increase in the transcript level at later time-points. These results suggest that CD63 might contribute to T. molitor immune response against various microbial pathogens.

Apolipophorin III (apoLp-III) is a well-known hemolymph protein having a functional role in lipid transport and immune response of insects. We cloned full-length cDNA encoding putative apoLp-III from larvae of the coleopteran beetle, Tenebrio molitor (TmapoLp-III), by identification of clones corresponding to the partial sequence of TmapoLp-III, subsequently followed with full length sequencing by a clone-by-clone primer walking method. The complete cDNA consists of 890 nucleotides, including an ORF encoding 196 amino acid residues. Excluding a putative signal peptide of the first 20 amino acid residues, the 176-residue mature apoLp-III has a calculated molecular mass of 19,146 Da. Genomic sequence analysis with respect to its cDNA showed that TmapoLp-III was organized into four exons interrupted by three introns. Several immune-related transcription factor binding sites were discovered in the putative 5’-flanking region. BLAST and phylogenetic analysis reveals that TmapoLp-III has high sequence identity (88%) with Tribolium castaneum apoLp-III but shares little sequence homologies (<26%) with other apoLp-IIIs. Homology modeling of Tm apoLp-III shows a bundle of five amphipathic helices, including a short helix 3’. The ‘helix-short helix-helix’ motif was predicted to be implicated in lipid binding interactions, through reversible conformational changes and accommodating the hydrophobic residues to the exterior for stability. Highest level of TmapoLp-III mRNA was detected at late pupal stages, albeit it is expressed in the larval and adult stages at lower levels. The tissue specific expression of the transcripts showed significantly higher numbers in larval fat body and adult integument. In addition, TmapoLp-III mRNA was found to be highly up-regulated in late stages of L. monocytogenes or E. coli challenge. These results indicate that TmapoLp-III may play an important role in innate immune responses against bacterial pathogens in T. molitor.

We have identified novel ricin-type (R-type) lectin by sequencing of random clones from cDNA library of the coleopteran beetle, T.molitor. The cDNA sequence is comprised of 495 bp encoding a protein of 164 amino acid residues and shows 49% identity with galectin of Tribolium castaneum. Bioinformatics analysis shows that the amino acid residues from 35 to 162 belong to ricin-type β-trefoil structure. The transcript was significantly upregulated after early hours of injection with peptidoglycans derived from Gram (+) and Gram (-) bacteria, beta-1, 3 glucan from fungi and an intracellular pathogen, L. monocytogenes suggesting putative function in innate immunity.

Rice stripe virus disease (RSVD), one of the most serious disease of rice is mediated through the sucking by small brown planthopper, Laodalphax striatellus. So far, the studies have been mainly focused on the interaction between the host plant and the virus. In this study, for better comprehension of the interactions among the host plant, vector insect and plant-pathogenic virus, we investigated transcriptome of the vector insect and the differences between viruliferous and naïve L.striatellus. For this, naïve L. striatellus were collected from non-infected rice field and 50 L.striatellus of them were fed RSV-infected rice for 5 days. With the RSV-viruliferous and the naïve insects, we conducted Illumina RNA sequencing (Hiseq 2000) and obtained 175,243,488 and 146,031,348 reads from viruliferous and naïve L.striatellus, respectively. These reads were assembled into contigs and two transcriptome databases were generated. The transcriptome of naïve and RSV-viruliferous L. striatellus were campared to figure out up-regulated or down-regulated genes. These RSV-dependently regulated genes may have important function in the behavior of planthoppers or the transmission of RSV.

본 연구는 국내 일반 사양환경에서 ROSS 육용계의 성장능력을 조사․분석하고, 사료효율을 고려한 도계의 적정시기를 산출하며, 도체성적을 분석하여 ROSS종 육계의 국내 활용 가능성을 검증하고, 성장관련 유용유전자를 발굴하여 조기선발을 위한 육종 전략을 수립하는데 목적으로 실시하였다. ROSS308 육계는 부분육 생산에 탁월한 능력을 지닌 품종으로 국외에서는 일반적으로 42일령에 도계를 실시하지만 국내에서는 55일령까지는 도계시기를 연장해도 좋지만 그 이후에는 사료효율이 저하되기 때문에 55일령을 도계시기로 선정하는 것이 바람직하다. 일반성장능력은 일당증체량이 83.4±10.4g, 사료섭취량은 4.5±1.4g, 출하체중은 4,833.2±570.7g, 정강이 길이는 9.36±5.4㎜, 가슴중량은 1,088.6± 125.0g, 넓적다리 중량은 864.8±86.9g으로 조사되었다. 초기 성장에서 능력이 저하되는 개체들은 조기 도태를 유도하여 계군 전체의 성장 능력 및 도체 성적을 개선할 수 있을 것으로 사료되며 이를 위하여 4개의 후보유전자인 TGFBR1, TGFBR1, IGF2, POUF1 등을 활용하여 조기선발을 유도하는 것이 유용할 것으로 사료된다.

Uncovering enzyme (UCE), encoded by the human NAGPA, is a trans-Golgi enzyme that adds the mannose-6- phosphate recognition tag on lysosomal enzymes destined for the lysosome. Mutations in NAGPA are known to cause stuttering, a common speech disorder with unknown etiology. The human NAGPA gene is transcribed into two different forms, probably due to alternative splicing. One of them, known as a brain isoform, is lacking exon 8 (102-bp). We performed quantitative real-time PCR for the NAGPA brain and non-brain isoforms in a cDNA panel originating from 16 human tissues and 24 sub-brain regions. According to our findings, the relative quantity of the NAGPA brain isoform in the brain was 4.7 times more than that in the control cDNA, a pooled mixture of equal amounts of cDNAs from the 16 different tissues. Further analysis using the cDNA panel originating from 24 different sub-brain regions revealed that the cerebral cortex contained the largest amount of NAGPA brain isoform. Relative quantity in the cerebral cortex was 8.6 times more than that in the control cDNA (P=0.00004). The lowest quantity of this isoform was detected in cDNA from the pituitary gland. In conclusion, findings of the current study suggest that the cerebral cortex, expressing the highest quantity of the NAGPA brain isoform, might be the region associated with speech function.