Molecular Cloning, Expression and Genomic Structure of Glyceraldehyde-3-Phosphate Dehydrogenase from the Entomopathogenic fungus, Paecilomyces tenuipes Jocheon-1

Nam Sook Park,Han Seok Kang,Seon Ku Kim,Yong Gyun Kim,Byung Uuk Cho,Teak Soon Shin,Keun Ki Kim,Hyun Chul Park,Hong Joo Son,Hong Gu Lee,Sang Mong Lee 한국응용곤충학회 2011 한국응용곤충학회 학술대회논문집 Vol.2011 No.05

Presently, We have constructed an olig-d(T) primed directional cDNA library from the silkworm Dongchunghacho, an entomopathogenic fungus, of which species is belonging to Paecilomyces tenuipes Jocheon-1. To isolate and screen genes in the fungus, 626 expressed sequence tags(ESTs) were generated by a partial sequencing from the cDNA library. Paecilomyces tenuipes Jocheon-1 cDNA encoding the glyceraldehyde-3-phosphate dehydrogenase(Pt-GAPDH) of Paecilomyces tenuipes Jocheon-1 was cloned from the above cDNA library. The complete cDNA sequence of Pt-GAPDHis comprised of 1,014bp encoding 338 amino acid residues. The deduced protein sequence of Pt-GAPDH showed higher homology with Beauberia bassiana-GAPDH(93% amino acid identity). Hydropathy analysis revealed that Pt-GAPDH protein is hydrophilic. The major three amino acids in its composition of amino acid residues were alanine(11.54%), valine(9.47%) and glycine(8.88%). The cDNA encoding Pt-GAPDH was expressed as a 37 kDa polypeptide in baculovirus-infected insect Sf9 cells. The Pt-GAPDH gene of Paecilomyces tenuipes entomopathogenic fungus consisted of three exons and two introns coding for 338 amino acid residues, and the genomic DNA length of the gene spans 1302bp. The accession number of the gene in GenBank are GU997099 for Pt-GAPDH cDNA and GU997102 for Pt-GAPDH genomic DNA.

cDNA Cloning and Genomic Structure of Glyceraldehyde-3-Phosphate Dehydrogenase from the Entomopathogenic fungus, Paecilomyces tenuipes-Jocheon

Nam Sook Park,Han Seok Kang,Seon Ku Kim,Yong Gyun Kim,Byung Uuk Cho,Teak Soon Shin,Keun Ki Kim,Hyun Chul Park,Hong Joo Son,Hong Gu Lee,Sang Mong Lee 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.05

Fungi belonging to the Paecilomyces spp. have recently been used as food and herbal medicines in Korea and are greatly popular as commercially available powdered supplement or dried fruiting body. Despite this acceptance and its use, little is known of the genes related to its reactive agents. Presently, We have constructed an olig-d(T) primed directional cDNA library from the silkworm Dongchunghacho, an entomopathogenic fungus, of which species is belonging to Paecilomyces spp. based on the previous identification of ITS1 and ITS2 at the molecular level and collected from Jocheon Miryang, Korea. To isolate and screen genes in the fungus, 626 expressed sequence tags(ESTs) were generated by a partial sequencing from the cDNA library. cDNA encoding the glyceraldehyde-3-phosphate dehydrogenase(Pt-GAPDH) of Paecilomyces tenuipes- Jocheon was cloned from the above cDNA library. The complete cDNA sequence of Pt-GAPDH is comprised of 1,014bp encoding 338 amino acid residues. The deduced protein sequence of Pt-GAPDH showed higher homology with Beauberia bassiana-GAPDH(93% amino acid identity). Hydropathy analysis revealed that Pt-GAPDH protein is hydrophilic. The major three amino acids in its composition of amino acid residues were alanine(11.54%), valine(9.47%) and glycine(8.88%). The Pt-GAPDH gene of Paecilomyces tenuipes entomopathogenic fungus consisted of three exons and two introns coding for 338 amino acid residues, and the genomic DNA length of the gene spans 1302bp. The accession number of the gene in GenBank are GU997099 for Pt-GAPDH cDNA and GU997102 for Pt-GAPDH genomic DNA. More investigation works including gene expression, immunological analysis etc. will be carried continuously without hesitation after this presentation.

돈분뇨의 토양처리시 이온 성분의 용탈 특성

김태헌,류성필,김성수,오윤근,허철구 한국환경과학회 2003 한국환경과학회지 Vol.12 No.3

An agricultural land application of swine slurry is one of the best management practices in Jeju island whose ground water must be protected. So as to study the effect of appling swine slurry on ground water or aquifer, incubation-leaching technique was used by assuming the incubating period of 1, 2, 4, 8, 16, or 32 days, and application rate of 3200.0㎎T-P/ℓ, 820.0㎎T-P/ℓ, and 1887.0㎎K^+/ℓ in swine slurry. The leachates were collected from the soil columns(PVC 30㎝L×5.5㎝D) packed 15㎝ in depth with Gangjeong soil series by washing with 100mL distilled water. The leached components were measured by using Ion chromatography for Cl^-, NO_3-N, F^-,Br^-, Na^+, K^+, Ca^2+, and Mg^2+, atomic absorption spectrophotometry for Fe and Mn, and UV-Vis spectrophotometry for T-N and T-P. Application of swine slurry in naked soil could influence on the ground water or aquifer by increasing nitrate-nitrogen in leachate with time, or leaching the cations present in soils in accompany with anions because of H^+ produced in nitrification. Therefore, careful consideration should be taken about what amount, when, where, and how for protecting ground water system.

GIS와 RS를 이용한 생태지도 작성기법에 관한 기초연구

이기철,이원화,윤해순,남춘희,김구연,김승환,서상현 한국지리정보학회 2004 한국지리정보학회지 Vol.7 No.3

본 연구는 낙동강 하구 일대의 생태계 변화와 생태 관련자료를 GIS DB로 구축하고 생태지도를 작성하였다. 생태지도 작성을 위해 1984년 11월 21일, 1997년 5월 17일에 촬영된 공간해상도 30m의 Landsat TM 위성영상과 국립지리원에서 발행된 1:25,000 수치지형도, 부산시에서 조사한 생물현황 자료를 바탕으로 DB를 구축하였다. 생태지도를 작성하기 위해, 첫째, 낙동강 하구 생태계 조사 보고서와 현재까지 진행된 생태지도에 대한 문헌 연구, 둘째, 시계열적 토지피복분류도 제작, 셋째, 동·식물상, 수질 등 생태계 항목별 BB 구축과 3단계 방식에 의한 생태계 평가, 최종적으로는 이러한 분석 결과를 바탕으로 생태계 관리를 위한 생태지도를 작성하였다. This study developed an ecological mapping technique with GIS database using the analyses of existing ecological survey reports and the change detection on the Nakdong river estuary. The data which are used to establish GIS DB include 2 Landsat TM images on Nov. 31, 1984 and May 17, 1997, 1:25, 000 topographical maps established by National Geography Institution and various ecological survey reports published by Busan metropolitan city government. The details for producing ecological map are as follows. At first, the current methods of ecomapping efforts and previous ecological surveys of Nakdong river estuary were carefully examined. Secondly, the land cover maps were created from the classified Landsat images of 1984 and 1997 for the spatiotemporal ecosystem analysis. Thirdly, the ecosystem was evaluated by using GIS ecological database based on the criteria of botany, zoology and water quality etc. Each criteria was reclassified into 3 stages which describe the overall quality of ecological condition. At last, the comprehensive ecological map was suggested as a prototype of ecosystem assesment and management tool with the discussion of further study. The findings of this study would be a milestone for preserving and managing the ecosystem.

외상성 뇌경막외 및 뇌경막하 혈종에서 혈종제거술 후 발생한 원발 병소 반대측 혈종에 대한 임상적 고찰

전은오,이상구,전영일,유찬종,김영보,박철완 최신의학사 2003 最新醫學 Vol.46 No.9-10

Pollen Viability of Common Buckwheat (Fagopyrum esculentum Moench.) in in vitro Germination and its Application to Male Gametophyte Isolation

Woo, Sun-HeeㆍKeun-Yook ChungㆍHeung-Tae, KimㆍYong-Gu, ChoㆍHong-Sig, KimㆍBeom-Heon, Songㆍ Chul-Won, Lee and Seung-Keun Jong 忠北大學校 農業科學硏究所 2004 農業科學硏究 Vol.21 No.-

-본문참고-

Morphological Traits and Cytogenetic Analysis of Intergeneric Hybrid between Neofinetia falcata and Sedirea japonicum

Chul Gu Been,Dong Gyu Kim,Sun Bin Kang,Jin Ki Kim,Byeong Jeong Lee,Hyun Yeol Shin,Kang Kwun Kim,Ki-Byung Lim 한국화훼산업육성협회 2014 화훼연구 Vol.22 No.4

The currently cultivated varieties of moth orchid flowers have weak or no fragrance and the plant requires high temperatures for cultivation. In order to develop a new variety of orchid that is psychrophilic and fragrant, intergeneric cross between wild wind orchids and moth orchids was performed. In 2009, To obtain intergeneric hybrids from Sedirea japonicum and Neofinetia falcata with moth orchids, artificial crosses with a total of 160 combinations were performed. Most of the cross combinations failed due to crossincompatibility after intergeneric crosses, with no pod formation, premature pod dropping after pod formation, and pod formation that did not produce seeds. From among these, the crosses that formed normal seeds and germinated to produce viable plants included Doritis pulcherrima × S. japonicum, N. falcata × D. pulcherrima, and N. falcata × P. equestris. From the hybrid specimens obtained through these crosses, 2 superior lines (819-3B and K9256) were selected based on their floral morphology, number of flowers, hardiness, and fragrance. Comparative analyses of the morphological and chromosomal traits were performed between the selected hybrid specimens and their parents. The flower and inflorescence characteristics of the selected hybrids exhibited intermediary traits of both the parents; however, more traits from the moth orchids were inherited. For pollinia traits, both 819-3B and K9256 specimens exhibited intermediate forms of their parents in terms of their shape and size. In the case of 819-3B specimen, a high degree of trait similarity with that of D. pulcherrima was observed; whereas, K9256 specimen showed lobes similar to that of P. equestris. The ploidy of moth orchid, S. japonicum, and N. falcata used for crosses was diploid, as determined by microscopic examination and ploidy analysis. The microscopic examination of chromosomes from the intergeneric hybrid pollens revealed mutation of the chromosome count among different specimens as well as irregular chromosome separation. For the production of next-generation progeny using the obtained hybrids, back-cross was performed; however, most of the progeny obtained were sterile and, among the 12 back-crosses, pod formation was noted in S. japonicum × 819-3B and 819-3B × S. japonicum crosses only.

당뇨 유도 백서 경골에 매식한 티타늄 임플란트 주위에 지방조직 유래 줄기세포 적용 시 골치유에 미치는 영향

김민구(Min-Gu Kim),정인교(In-Kyo Jung),신상훈(Sang-Hun Shin),김철훈(Chul-Hoon Kim),김복주(Bok-Joo Kim),김정한(Jung-Han Kim),황영섭(Young-Sup Hwang),정유진(Eu-Gene Jung),김진우(Jin-Woo Kim),김욱규(Uk-Kyu Kim) 대한구강악안면외과학회 2010 대한구강악안면외과학회지 Vol.36 No.5

Introduction: Diabetes mellitus, as a major health problem for the elderly has been shown to alter the properties of the bone and impair bone healing around a titanium implant in both humans and animals. The aim of this study was to examine the effect of adipose-derived stem cells on the healing process around a titanium implant in streptozotocin-induced diabetic rats. Materials and Methods: Thirteen rats were divided into two groups: adipose-derived stem cells injected group and a control group. A titanium screw implant (diameter: 2.0 mm, length: 3.5 mm) was placed into both tibia of 13 rats: 13 right tibia as the control group and 13 left tibia as the experimental group. The rats were sacrificed at different intervals (1, 2, and 4 weeks) after implantation for histopathology observations and immunohistochemistric analysis. Results: The histopathological findings revealed earlier new formed bone in the experimental group than the control group. In particular, at 1 week after implantation, the experimental group showed more newly formed bone and collagen around the implant than the control group. In immunohistochemistric analysis, osteoprotegerin (OPG) expression in the experimental group increased early compared to that of the control group until 2 weeks after implantation. However, after 2 weeks, OPG expression in the experimental group was similar to OPG expression in the control group. The receptor activator of nuclear factor κB ligand (RANKL) expression in the experimental group increased early compared to that of the control group, and then decreased at 2 weeks. After 2 weeks, the level of RANKL expression was similar in both groups. Conclusion: These results suggest that adipose-derived stem cells in implantation can promote bone healing around titanium, particularly in diabetes mellitus induced animals.

Brain somatic mutations in MTOR cause focal cortical dysplasia type II leading to intractable epilepsy

Lim, Jae Seok,Kim, Woo-il,Kang, Hoon-Chul,Kim, Se Hoon,Park, Ah Hyung,Park, Eun Kyung,Cho, Young-Wook,Kim, Sangwoo,Kim, Ho Min,Kim, Jeong A,Kim, Junho,Rhee, Hwanseok,Kang, Seok-Gu,Kim, Heung Dong,Kim, Nature Publishing Group 2015 Nature medicine Vol. No.

Focal cortical dysplasia type II (FCDII) is a sporadic developmental malformation of the cerebral cortex characterized by dysmorphic neurons, dyslamination and medically refractory epilepsy. It has been hypothesized that FCD is caused by somatic mutations in affected regions. Here, we used deep whole-exome sequencing (read depth, 412–668×) validated by site-specific amplicon sequencing (100–347,499×) in paired brain-blood DNA from four subjects with FCDII and uncovered a de novo brain somatic mutation, mechanistic target of rapamycin (MTOR) c.7280T>C (p.Leu2427Pro) in two subjects. Deep sequencing of the MTOR gene in an additional 73 subjects with FCDII using hybrid capture and PCR amplicon sequencing identified eight different somatic missense mutations found in multiple brain tissue samples of ten subjects. The identified mutations accounted for 15.6% of all subjects with FCDII studied (12 of 77). The identified mutations induced the hyperactivation of mTOR kinase. Focal cortical expression of mutant MTOR by in utero electroporation in mice was sufficient to disrupt neuronal migration and cause spontaneous seizures and cytomegalic neurons. Inhibition of mTOR with rapamycin suppressed cytomegalic neurons and epileptic seizures. This study provides, to our knowledge, the first evidence that brain somatic activating mutations in MTOR cause FCD and identifies mTOR as a treatment target for intractable epilepsy in FCD.

GSTM1, GSTT1, and GSTP1 Gene Polymorphisms Modify the Effect of Smoking on Serum Immunoglobulin E Level

Kim, Jin-Hee,Kim, Yong-Kyu,Park, Shin-Gu,Choi, Ji-Ho,Kim, Cheol-Woo,Lee, Kwan-Hee,Ha, Eun-Hee,Hong, Yun-Chul The Korean Society of Toxicogenomics and Toxicopro 2006 Molecular & cellular toxicology Vol.2 No.1

Immunoglobulin E (IgE) plays an important role in the development of allergic disorders including asthma. Cigarette smoking was reported to elevate serum IgE level and air pollutants such as $NO_{2}$ have been reported to modulate the immune system including inflammation. Moreover, genetic polymorphisms of glutathione S-transferases (GSTs) were reported to affect inflammatory diseases including asthma. Therefore, in the present study we tried to investigate whether tobacco smoke or $NO_{2}$ exposure increases the level of IgE and the GST gene polymorphisms are associated with change of IgE level due to tobacco smoke or $NO_{2}$ exposure. We measured urinary cotinine, personal $NO_{2}$ exposure, and serum IgE levels in 300 healthy university students without allergic disorders. Allelic loss of the GSTM1 and GSTT1 and the GSTP1 (lle105Val) polymorphism were determined by PCR and RFLP. Total serum IgE levels were significantly different according to urinary cotinine levels (P=0.046), while $NO_{2}$ passive dosimeter level and genetic polymorphisms of three GSTs were not associated with total IgE level. Moreover, subjects with cotinine $500\;{\mu}g/g$ creatinine or more showed the highest level of total IgE when they had null type of GSTM1, null type of GSTT1, or variant type of GSTP1 (P<0.05). When we considered IgE level according to urinary cotinine levels in strata with the combinations of GSTM1, GSTT1, and GSTP1 genetic polymorphisms, the subjects with GSTM1 null, GSTT1 null, and GSTP1 variant types showed the largest difference between IgE levels of subpopulations according to cotinine levels (P=0.030). However, there was no significant difference between IgE levels of subpopulations according to $NO_{2}$ passive dosimeter levels in any group with combinations of GSTM1, GSTT1, and GSTP1 polymorphisms. This result suggests that smoking increases allergic response measured as IgE level and combinations of the GSTM1, GSTT1, and GSTP1 polymorph isms modify the effect of smoking on serum IgE level.