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.

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.

위암에서 Galectin-3 단백질 발현에 관한 연구

주홍구,우희종 大韓免疫學會 1996 大韓免疫學會誌 Vol.18 No.4

The expression of galectin-3 protein in four different gastric cancer cell lines and lymph node tissues of patients with gastric carcinoma was studied by Western blotting, immundhistochenustry and flow cytometry analysis. Western blot analysis of galectin-3 using anti-galectin-3 , mAb indicated that a 29 kDa protein was expressed in 3 gastric cancer cell lines, AGS, KATO BI , and SNU16, but not in SNU1 which was a non-adherent cell line. Interestingly, SNU16 cells secreted galectin-3 into the culture media. Flow cytometry analysis revealed that NIH3T3, AGS, and KATO III cells expressed the galectin-3 on the cell surface. The level a))' galectin-3 expression was highest in AGS (91.5%), high in KATO M (69.5%), and low in NIH3T3 (55.8%). The gastric cancer cells metastasized into .lymph node tissues were heavily stained whereas normal lymphocytes were not stained. This study suggested that the expression of galectin-3 and the metastasis of gastric cancer cells might be correlated.

활성형 Vitamin D₃ 전환능을 소유한 방선균에서 P-450 Hydroxylase유전자의 클로닝 전략

서주원,현창구,김정미,김승영,홍순광 명지대학교 자연과학연구소 1998 자연과학논문집 Vol.17 No.-

비타민 D₃는 포유동물이나 새의 간에서 25-hydroxyvitamin D₃[25(OH)D₃]로 전환되는데, 이 hydroxylation은 P-450 hydroxylase에 의해 촉매된다고 알려져있다. 본 연구에서는 활성형 비타민 D₃전환능을 소유한 방선균에서P-450 hydroxylase효소를 신속하게 분리가능한 PCR방법을 보고하고자 한다. Primer는 방선균에서 보고된 P-450 hydroxylase들 사이의 아미노산 서열의비교를 통해 산소결합부위와 heme-ligand pocket 주의에서 제작되었고,5종의 방선균에서 효과적으로 PCR증폭산물을 획득할 수 있었다. 증폭산물을 획득할 수 있었다.증폭 산물들의 아미노산서열을 분석한 결과 기존에 밝혀진 많은P-450 hydroxylase유전자들과 상동성을 보이고 있었으며, 특히 Streptomyces griseolus에서 분리되어 sulfonylurea 등의 제초제의 분해능력을 가진 SuaC, SubC 효소 등과 높은 아미노산 상동성을 지니고 있었다. 이러한 PCR전략은 방선균에서 유용한 물질의 대사에 관여하는 P-450 hydroxylase효소의 클로닝에 도움을 줄 것이라 사료된다 In mammals and birds, vitamin D₃is converted to 25-hydroxyvitamin D₃[25(OH)D₃] and then to 1α25-dihydroxyvitamin D₃[25(OH)₂D₃]in the liver and kideny, respectively these hydroxylating reactions are known to be catalyzed by P-450 hydroxylases. In this paper we report a PCR method which can be used for the repid amplification of DNA fragments for _450 hydroxylase from actinomycetes which can convert to active from of vitamin D₃. Primers were designed based on several regions sharing strong similarities in amino acid sequence of P-450 hydroxylases from a variety of actinomycetes, primarily in the regions of an oxygen binding site and a heme ligand pocket. These primers were used to amplify DNA fragments from five different actinomycetes. The deduced amino acid sequences of the isolated fragments revealed significant similarities to known P-450 hydroxylase including the product of the suaC or subC genes from Streptomyces griseolus that is capable of metabolizing a number of sulfonylurea herbicides and to the product of the P450sca2 from S. carbophilus that produces a specific HMG-CoA reductase inhibitor. This method will be helpful for the researchers in cloning the genes for P-450 hydroxylase involved in the biosynthesis of useful compounds.

유압관로내의 동특성을 이용한 비정상 유량계측에 관한 연구

이홍구,윤석주,박성규,박길문 朝鮮大學校 機械技術硏究所 1998 機械技術硏究 Vol.1 No.1

This paper describes an approach for estimating the unsteady flowrate through a hydraulic pipe lines and a components in real time. Concerning the dynamic characteristics of a hydraulic pipe lines, experimental research from the view point of focusing on the unsteady flowrate through a pipe is insufficient because of the lack of adequate flowmeters. In this experiment, a new instantaneous flowrate measurement system making use of a sharp-edged cylindrical choke and LDV has been proposed and developed by the authors. The unsteady flowrate is obtained from the measurements center-line velocity of a circular pipe using LDV and the pressure difference using a sharp-edged cylindrical choke, and experiments on the frequency characteristics are carried out. The results show a good agreement between the estimated and directly measured flowrate waveforms illustrates the validity of the method proposed here.

일부 직장인의 정기 건강진단 실시 결과에 관한 고찰

남택승,홍종관,박종안,한구웅,김주자 순천향대학교 1987 논문집 Vol.10 No.2

서울, 천안, 구미에 각각 위치한 순천향대학 부속병원에서 서울, 천안, 구미 및 인근 군단위 지역의 공무원 및 사립학교 교직원을 대상으로 1986년 4월부터 동년 7월까지 집단건강진단을 실시하고 설문지를 배부, 조사하여 분석한 결과 다음과 같은 성적을 얻었다. 1. 1차 검사결과 유소견율은 수축기혈압 1.1%(남자 1.4%, 여자 0.2%), 이완기혈압 6.4%(남자 8.0%, 여자 1.5%)이었다. 2. 공복시혈당은 전대상자의 3.3%(남자 3.8%, 여자 1.2%)가 고혈당의 유소견율을 나타내었으며 뇨당은 1.3%(남자 1.6%, 여자 0.5%)의 유소견율을 보였다. 3. 순환기계질환에 관한 2차 검사유소견율은 수축기혈압 24.1%, 이완기혈압 74.9%.안저검사 41.1%, 심전도 검사 36.1%. 혈중 트리글리세라이드 검사 36.1%이었다. 4. 혈압의 연도별 변화는 '86년도의 혈압 평균치는 수축기혈압이 남자 122.7±15.7mmHg, 여자 113.0±13.1mmHg로 '82. '84년도에 비해 유의한 차가 있는 것으로 나타났으며(남 P<0.01, 여 P<0.05) 이완기혈압은 남자 80.9±15.1mmHg, 여자 73.8±23.9mmHg로 남자에 있어서만 (P<0.05). 연도별로 유의한 차가 있는 것으로 나타났다. 5. 혈압의 개인별 경시적 변화를 보면 수축기혈압의 경우 전체적으로 99.8%가 계속 정상으로 유지되고 1.2%는 고혈압 상태로 계속 유지되고 있는 것으로 나타났으며 이완기혈압의 경우는 전체적으로 92.8%가 계속 정상, 7.1%가 계속 고혈압으로 나타났다. 6. 혈중 콜레스테롤 평균치의 연도별 변화는 통계적으로 유의하게 증가되었다.(P<0.01). 7. 혈압과 관련요소들에 대한 상관관계에서 상관성이 있는 것은 수축기혈압의 경우 남자에 있어서 비만도, 뇨단백, 혈중 크레아티닌 및 콜레스테롤, 안저검사, 심전도검사, 혈중 트리글리세라이드이었고 여자에 있어서는 Stout지수, Rohrer지수, 뇨단백, 혈중 콜레스테롤, 안저검사, 심전도검사, 혈중 트리글리세라이드이었다. 이완기혈압의 경우 남자에 있어서는 뇨단백, 혈중 콜레스테롤, 안저검사, 심전도 검사, 혈중 트리글리세라이드이었고 여자에 있어서는 Stout지수, Rohrer지수, 뇨단백, 혈중 크레아티닌, 심전도 검사, 혈중 트리글리세라이드이었다. 8. 당뇨질환에 관한 2차 검사 유소견율은 공복시 혈당이 전체 51.8%, 식후 2시간 혈당치가 전체 41.2%이었다. 9. 공복시 혈당 평균치의 연도별 변화에서 '86년도 혈당 평균치는 남자 92.5±20.4㎎%, 여자 87.6±15.9㎎%이었고 연도별 증가는 통계적으로 유의하였다(P<0.01). 10. 혈당의 개인별 경시적 변화는 전체적으로 99.6%가 계속 정상, 2.32%가 계속 고혈당이었고 고혈당에서 정상으로, 정상에서 고혈당으로 된 경우는 각각 0.02%, 0.05%였다. 11. 혈당과 관련요소들에 대한 상관관계에서 상관성이 있는 것은 남녀 모두에서 Stout지수, Rohrer지수, 뇨당, 식후 2시간 혈당치이었다. 12. 건강행태에 관한 설문조사에서 자신의 건강상태에 대한 인식, 흡연, 음주, 식사에 대한 항목이 정상군과 유소견군간에 유의한 차가 없는 것으로 나타났다. Health Insurance Corporation which covers government officials and privated school faculties has held periodic health examination for the insured every other years since 1982 under the Insurance Law. The purpose is to detect diseases and trea diseases at their early stage. Three Soonchunhyang University Affiliated Hospitals located each in Seoul, Chunan, and Gumi city are committed to carry out the health examination. There are two steps in examination. The first step is for all insured, the second step is for the people whose results come out abnormal in the first step examination. Each item of the tests has been predetermined by the Corporation and most of them are to detect many chronic diseases e.g. hypertension, diabetes mellitus, liver disease, pulmonary Tbc., renal disease, and venereal disease. In spite of increasing concern regarding these diseases recently, we are lack of even basic statistics such as prevalence rate. Authors collected health examination results from three hospitals and analysed them.

Responses of Plasma IGF-1, IGFBPs and Hepatic GH Receptor to Growth Hormone Releasing Peptides (GHRP)-2 Administration and Energy Level in Wethers

Hong Gu Lee(이홍구),Young Cheng Jin(김영성),Hisashi Hidari,Yun Jaie Choi(최윤재),Seon Ku Kim(김선구),Teak Soon Shin(신택순),Byung Uuk Cho(조병욱),Yong Gyun Kim(김용균),Keun Ki Kim(김근기),Hong Joo Son(손홍주),Sang Mong Lee(이상몽) 한국생명과학회 2008 생명과학회지 Vol.18 No.7

본 연구는 정상으로 단백질을 급여한 거세면양에 있어서 에너지 첨가가 GHRP-2투여에 대한 혈장 IGF-1 및 IGFBPs에 대한 반응과, 고에너지 급여에 따른 GHRP-2투여가 hepatic GH 수용체에 미치는 영향을 검증하기 위하여 실시하였다. 시험 결과 HENP (CP 0.34 ㎏, TDN 1.83 ㎏/day DM intake)처리기간 동안 혈장 IGF-1 과 39-42kDa IGFBP-3수준은 LENP (CP 0.32 ㎏, TDN 0.87 ㎏/day DM intake)기간에 비하여 높게 나타났으나 (P<0.05), 혈장 34 kDa IGFBP-2와 24 kDa IGFBP-4는 영양처리에 의해 영향을 받지 않았다. 각 영양처리 기간동안 GHRP-2 (12.5 μg/㎏ body weight/day)투여는 혈장 GH 반응이 촉진되었으며(P<0.05), 혈장 GH 평균 함량과 AUC증가에 있어서는 LENP처리 기간에 비하여 HENP처리기간에서 유의적으로 높게 나타났다(P<0.01). 특히 HENP에서 7일간 GHRP-2투여에 의한 일중 혈장 IGF-1 변화양상을 조사한 결과 투여 2, 6 및 7일에서 뚜렷한 증가양상을 보였다(P<0.05). 이에 반하여 LENP에서는 오직 투여 3일째에서 Saline구에 비하여 유의적인 증가를 확인하였다(P<0.05). IGFBPs의 ligand blotting 결과 HENP구에서 혈장 39-43 kDa IGFBP-3의 수준의 증가가 투여 6일과 7일에서 관찰되었으나 혈장 IGFBP-2수준은 두 영양처리시기에서 유의적인 차이를 관찰하지 못했다. 아울러 HENP구에 있어서 간세포막에 <SUP>125</SUP>I-oGH의 결합력을 측정한 결과 GHRP-2투여에 의한 영향은 관찰되지 않았다. 이와 같은 결과는 거세면양에 있어서 단백질과 에너지 사이의 영양적 균형은 내인성 GH/IGF-1 axis는 물론 혈장 IGFBP-3수준의 변화에 영향을 미치고 있음을 시사한다. The purpose of this study was to determine the effect of energy supplement on responses of plasma insulin-like growth factor (IGF)-1 and IGF binding proteins (IGFBPs) to growth hormone-releasing peptide-2 (GHRP-2) administration in normal protein-fed wethers, and to observe the effect of GHRP-2 treatment on hepatic growth hormone (GH) receptor in well-fed wethers. Plasma IGF-1 and 39-42 kDa IGFBP-3 during the HENP (CP, crude protein 0.34 and TDN, total digestible nutrients 1.83 ㎏/day DM, dry matter intake) treatment period were higher than in the LENP (CP 0.32 ㎏ and TDN 0.87 ㎏/day DM intake) period (P<0.05). The response of GH was stimulated by GHRP-2 (12.5 ㎍/㎏ body weight/day) administration during both of the feed treatment periods (P<0.05). The area under curve (AUC) increment and average concentration of GH (0-180 min) with GHRP-2 administration was higher during HENP treatment than LENP treatment (P<0.01). During the HENP treatment period from day 1 to day 7 of twice daily GHRP-2 treatment, the plasma IGF-1 increment was increased on days 2, 6 and 7 of GHRP-2 administration (P<0.05). On the basis of ligand blotting, the proportions of plasma 39-43 kDa IGFBP-3 during the HENP treatment period only showed a significant difference on days 6 and 7 with GHRP-2 administration. No significant difference in the specific binding of 125I-labeled oGH to hepatic membranes was detected between the saline and GHRP-2 treatments of the HENP-fed wethers. These results suggest that the nutritional balance between energy and protein may affect the endogenous GH / IGF-1 axis as well as plasma IGFBP-3 levels.

Radioprotective effects of an acidic polysaccharide of Panaxginseng on bone marrow cells

Hong-Gu Joo,Hyun-Ji Kim,Mi-Hyoung Kim,Yun-Young Byon,Jae Woo Park,Youngheun Jee 대한수의학회 2007 JOURNAL OF VETERINARY SCIENCE Vol.8 No.1

An acidic polysaccharide of Panax ginseng (APG), so called ginsan is known to have important immunomodulatory activities. It was recently reported that APG has radioprotective effects in mice but the detailed mechanism was not fully elucidated. This study examined the effects of APG on bone marrow cells (BMs). The phenotypical and functional changes in APG-treated BMs after gamma radiation were studied. The benefit of APG on BMs damaged by gamma radiation was determined by measuring the cell viability. Using 2 different assays, a pretreatment with APG significantly increased the viability of BMs against gamma radiation. APG-treated BMs had a significantly higher amount of IL-12, which is a major cytokine for immune responses, compared with the medium-treated BMs. The expression of MHC class II molecules of APG-treated BMs was also increased, and APG-treated BMs showed significantly higher levels of allogeneic CD4+ T lymphocyte proliferation. Furthermore, APG-treated mice had a larger number of BMs after gamma radiation than the control mice, and the BMs of APG-treated mice were successfully cultured into dendritic cells, which are the representative antigenpresenting cells. Overall, this study shows that APG alters the phenotype of BMs, increases the viability and alloreactivity of BMs after gamma radiation both in vitro and in vivo. Therefore, APG may be a good candidate radioprotective agent for BMs.

A Novel Function of Interferon Regulatory Factor-1.

Hong-gu Joo 대한수의학회 2003 대한수의학회 학술대회발표집 Vol.2003 No.-

Radioprotective activity and stimulatory effects of an acidic polysaccharide of Panax ginseng on bone marrow and immune cells

Hong-Gu Joo(주홍구) 고려인삼학회 2011 고려인삼학회 학술대회 Vol.2011 No.4