• Title/Summary/Keyword: Gene Identification

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MicroRNA-23b is a Potential Tumor Suppressor in Diffuse Large B-cell Lymphoma (미만성 거대 B 세포 림프종(DLBCL)에서 microRNA-23b의 잠재적 종양 억제자로서의 효과)

  • Nam, Jehyun;Kim, Eunkyung;Kim, Jinyoung;Jeong, Dawoom;Kim, Donguk;Kwak, Bomi;Kim, Sang-Woo
    • Journal of Life Science
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    • v.27 no.2
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    • pp.149-154
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    • 2017
  • Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-hodgkin lymphoma. Advances in the chemotherapeutic treatment of this disease have improved the outcomes of DLBCL; nonetheless, many patients still die of DLBCL, and therefore, a better understanding of this disease and identification of novel therapeutic targets are urgently required. In a recent gene expression profiling study, PDE (phosphodiesterase) 4B was found to be overexpressed in chemotherapy-resistant tumors. The major function of PDE4B is to inactivate the second messenger cyclic 3',5' monophosphate (cAMP) by catalyzing the hydrolysis of cAMP to 5'AMP. It is known that cAMP induces cell cycle arrest and/or apoptosis in B cells, and PDE4B abolishes cAMP's effect on B cells. However, the mechanism by which PDE4B is overexpressed remains unclear. Here, we show that the aberrant expression of miRNA may be associated with the overexpression of this gene. The PDE4B 3' untranslated region (UTR) has three functional binding sites of miR-23b, as confirmed by luciferase reporter assays. Interestingly, miR-23b-binding sites were evolutionarily conserved from humans to lizards, implying the critical role of PDE4B-miR-23b interaction in cellular physiology. The ectopic expression of miR-2 3b repressed PDE4B mRNA levels and enhanced intracellular cAMP concentrations. Additionally, miR-23b expression inhibited cell proliferation and survival of DLBCL cells only in the presence of forskolin, an activator of adenylyl cyclase, suggesting that miR-23b's effect is via the downregulation of PDE4B. These results together suggest that miR-23b could be a therapeutic target for overcoming drug resistance by repressing PDE4B in DLBCL.

ISOLATION AND IDENTIFICATION OF BACTERIA FROM THE ROOT CANAL OF THE TEETH DIAGNOSED AS THE ACUTE PULPITIS AND ACUTE PERIAPICAL ABSCESS (급성 치수염 및 급성 치근단 농양의 치근관으로부터의 세균 분리 및 동정)

  • Lee, Yeon-Jae;Kim, Mi-Kwang;Hwang, Ho-Keel;Kook, Joong-Ki
    • Restorative Dentistry and Endodontics
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    • v.30 no.5
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    • pp.409-422
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    • 2005
  • The aim of this study was to identify the bacteria isolated from acute endodontic lesions by cell culture and 16S rDNA sequencing. The necrotic pulpal tissue was collected from 17 infected root canals, which were diagnosed as being either an acute pulpitis or acute periapical abscess. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ul of 1 XPBS. The sample solution was briefly mixed and plated onto a BHI-agar plate containing $5\%$ sheep blood. The agar plates were incubated in a $37^{\circ}C$ anaerobic chamber for 7 days. The bacteria growing on the agar plate were identified by 16S rRNA coding gene (rDNA) cloning and sequencing at the species level. Among the 71 colonies grown on the agar plates, 56 strains survived and were identified. In dental caries involving the root canals, Streptococcus spp. were mainly isolated. Actinomyces, Clostridia, Bacteroides and Fusobacteria were isolated in the periapical lesion without dental caries. Interestingly, two new Actinomyces spp. (ChDC B639 and ChDC B631) were isolated in this study. These results showed that there was diversity among the species in endodontic lesions, This suggests that an endodontic infection is a mixed infection with a polymicrobial etiology. These results may offer the bacterial strains for pathogenesis studies related to an endodontic infection.

Analysis of Genetic Polymorphism by Bloodtyping in Jeju Horse (혈액형에 의한 제주말의 유전적 다형성 분석)

  • Cho Gil-Jae
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.972-978
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    • 2005
  • The present study was carried out to investigate the blood markers of Jeju horses. The redcell cypes (blood groups) and blood protein types (biochemical polymorphisms) were tested from 102 Jeju horses by serological and electrophoretc procedure, and their phenotypes and gene frequencies were estimated. The blood group and biochemical polymorphism phenotypes observed with high frequency were $A^{af}\;(27.45\%$), $C^{a}\;(99.02\%$), $K^{-}\;(97.06\%$), $U^{a}\;(62.75\%$), $P^{b}\;(36.27\%$), $Q^{c}\;(47.06\%$), $D^{cgm/dghm}\;(13.73\%$), $D^{adn/cgm}\;(9.80\%$), $D^{ad/cgm}$\;(8.82\%$), $D^{dghm/dghm}(7.84\%$), $D^{cgm/cgm}(7.84\%$), $AL^{B}\;(48.04\%$), $GC^{F}\;(99.02\%$), $AlB^{K}\;(97.06\%$), $ES^{FI}\;(36.27\%$), $TF^{F2}\;(25.49\%$), $HB^{B1}\;(45.10\%$), and $PGD^{F}\;(86.27\%$) in Jeju horses, respectively. Alleles observed with high gene frequency were $A^{af}$ (0.3726), $A^{C}$ (0.2647), $C^{-}$ (0.5050), $K^{-}$ (0.9853), $U^{-}$ (0.6863), $P^{b}$ (0.4657), $Q^{c}$ (0.5294), $D^{cgm}$ (0.3039), $HB^{B1}$(0.6863), $PGD^{F}$ (0.9265), $AL^{B}$ (0.6912), $ALB^{K}$ (0.9852), $GC^{F}$ (0.9950), $ES^{I}$ (0.5000) and $TF^{F2}$ (0.4950) in Jeju horses, and sfecific alleles, $D^{cgm(f)}$ (0.0196), $HB^{A}$ (0.0147), $HB^{A2}$ (0.0196), $ES^{G}$ (0.0441), $ES^{H}$ (0.0098), $TF^{E}$TF'(0.0246), $TF^{H2}$ (0.0049) and $PGD^{D}$ (0.0098) were detected in Jeju horses. These preliminary results present basic information for detecting the genetic markers in Jeju horse. and developing a system for parentage verification and individuals identification in jeju horses.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
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    • 2015.05a
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    • pp.45-46
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    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

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Human coronavirus infection in hospitalized children with community-acquired pneumonia (입원한 폐렴 환아에서 코로나 바이러스 감염)

  • Chung, Ju-Young;Han, Tae Hee;Kim, Sang Woo;Koo, Ja Wook;Hwang, Eung-Soo
    • Pediatric Infection and Vaccine
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    • v.14 no.1
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    • pp.69-74
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    • 2007
  • Purpose : Human coronanviruses (hCovs) including hCoV-229E and hCoV-OC43 have been known as etiologic agents of the common colds and were regarded as clinically insignificant agents. However, recent identification of hCoV-NL63 and hCoV-HKU1 in children with lower respiratory tract infections has evoked the clinical concerns about their prevalence and the clinical significance of these hCoVs in children. This study was performed to investigate the prevalence of hCoVs in children with community-acquired pneumonia. Methods : From March 2006 to January 2007, nasopharyngeal specimens collected from children hospitalized with pneumonia, were tested for the presence of common respiratory viruses (respiratory syncytial virus, influenza A, influenza B, parainfluenza viruses, and adenovirus) using multiplex reverse transcriptase polymerase chain reaction (RT-PCR). Human metapneumovirus (hMPV) infection was excluded by nested RT-PCR using primers for the F-gene. To detect the different strains of hCoVs, nested RT-PCR assays specific for hCoVNL63, hCoV-OC43, hCoV-229E, and hCoV-HKU1 were performed. Results : Out of the 217 nasopharyngeal aspirate from children aged under 15 years, respiratory syncytial virus (RSV) was detected in 32 patients, hMPV in 18, human parainfluenza virus in 10, influenza virus A in 2, and adenovirus in 6. HCoVs were detected by RT-PCR in 8 (3.7%) of the 217 patients, hCoV-229E in 1, hCoV-NL63 in 3, and hCoVOC43 in 4 patients. HCoV-HKU1 was not detected in this study population. Conclusion : Recently identified hCoV-NL63 and hCoV-HKU1 seemed to have a little clinical significance in Korean children with severe or hospitalized community-acquired pneumonia.

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Identification of Differentially Expressed Genes Between Somatic Cell Nuclear Transfer and Normal Placenta in Cattle (소의 체세포핵이식태반과 정상태반간의 차등 발현 유전자 분석)

  • Yu, Seong-Lan;Jeong, Hang-Jin;Sang, Byung-Chan;Ryoo, Seung-Heui;Jung, Kie-Chul;Yoon, Jong-Taek;Seong, Hwan-Hoo;Jin, Dong-Il;Lee, Jun-Heon
    • Journal of Animal Science and Technology
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    • v.50 no.5
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    • pp.641-648
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    • 2008
  • There has been great success for making transgenic animals using somatic cell nuclear transfer(SCNT) up to this time. However, the success rates of the production of live transgenic animals are still very low. The current research has been carried out for delineation of differentially expressed genes between SCNT and normal placenta in cattle. In the present observations, high expression has been observed for CTSZ, LOC509426 and ELF1 genes in normal placenta. On the other hand, TIMP2, PAG1B, PAG-21, LOC782894, SERPINB6 and mKIAA2025 protein were highly expressed in SCNT placenta. Five genes, which were highly expressed in SCNT placenta, have been further investigated using semi-quantitative real-time PCR. The results were similar to that we observed using ACP. In the future, all genes affecting the SCNT and normal placenta have to be discovered and their networks will be fully investigated. The genes were identified in this study would be great help for identifying differential gene expressions in SCNT placenta.

Identification of Quantitative Trait Loci Associated with Anthracnose Resistance in Chili Pepper (Capsicum spp.) (고추 탄저병 저항성 관련 양적형질 유전자좌 분석)

  • Kim, Su;Kim, Ki-Taek;Kim, Dong-Hwi;Yang, Eun-Young;Cho, Myeong-Cheoul;Jamal, Arshad;Chae, Young;Pae, Do-Ham;Oh, Dae-Geun;Hwang, Ju-Kwang
    • Horticultural Science & Technology
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    • v.28 no.6
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    • pp.1014-1024
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    • 2010
  • Pepper ($Capsicum$ spp.) anthracnose caused by $Colletotrichum$ $acutatum$ is a destructive disease susceptible to areas where chili peppers are grown. $Capsicum$ $baccatum$ var. $pendulum$ (Cbp) is resistant to anthracnose and has actively been used for interspecific hybridization for the introgression of resistance gene(s) into cultivated chili peppers. The goals of this study were to determine the inheritance of resistance to anthracnose within $Capsicum$ $baccatum$ and to map quantitative trait loci (QTLs) for the anthracnose resistance. A genetic mapping population consisting of 126 $F_2$ plants derived from a cross between $Capsicum$ $baccatum$ var. $pendulum$ (resistant) and $Capsicum$ $baccatum$ 'Golden-aji' (susceptible) was used for linkage mapping. The linkage map was constructed with 52 SSRs, 175 AFLPs, and 100 SRAPs covering 1,896cM, with an average interval marker distance of 4.0cM. Based on this map, the number, location, and effect of QTLs for anthracnose resistance were studied using plants inoculated in the laboratory and field. A total of 19 quantitative trait loci (2 major QTLs and 16 minor QTLs) were detected. Two QTLs ($An8.1$, $An9.1$) showed 16.4% phenotypic variations for anthracnose resistance after wounding inoculation. In addition, five minor QTL loci ($An7.3$, $An7.4$, $An4.1$, $An3.1$, $An3.2$) showed a total of 60.73% phenotypic variations of anthracnose resistance in the field test. Several significant QTLs were also detected and their reproducibility was confirmed under different inoculation conditions. These QTLs are now being confirmed with different breeding populations. Markers tightly linked to the QTLs that are reliable under different environmental conditions will help to determine the success of marker-assisted selection for anthracnose -resistant breeding programs in chili pepper.

Current status, mechanism and control of herbicide resistant weeds in rice fields of Korea (한국 논에서 제초제 저항성잡초의 발생 현황, 메카니즘 및 방제)

  • Park, Tae Seon;Seong, Ki Yeong;Cho, Hyun Suk;Seo, Myung Chul;Kang, Hang Won;Park, Kee Woong
    • Korean Journal of Agricultural Science
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    • v.41 no.2
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    • pp.85-99
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    • 2014
  • Sulfonylurea (SU)-resistant weeds of eight annual weeds, Monochoria vaginalis, Scirpus juncoides and Cyperus difformis, etc., and four perennial weeds, Scirpus planiculmis, Sagittaria pigmaea, Eleocharis acicularis and Sagittaria trifolia as of 2013 since identification Monochoria korsakowii in the reclaimed rice field in 1998. And the resistant Echinochloa oryzoides to ACCase and ALS inhibitors has been confirmed in rice fields of the southern province, Korea in 2009. In the beginning, the M. vaginalis, S. juncoides and C. difformis of these SU-resistant weeds were rapidly and individually spreaded in different fields, however, these resistant weeds have been occurring simultaneously in the same filed recently. The resistant biotype by weed species demonstrated about 10-to 1,000-fold resistance, based on $GR_{50}$ values of the SU herbicides tested. And the resistant biotype of E. oryzoides to cyhalofop-butyl, pyriminobac-methyl, and penoxsulam was about 14, 8, and 11 times more resistant than the susceptible biotype base on $GR_{50}$ values. The products and applied area of SU-included herbicides have been increased rapidly, and have accounted for about 69% and 96% in Korea, respectively. In Korea, the main cause of SU-resistant weed is extensive use of these herbicides. The top ten herbicides by applied area were composed of all SU-included herbicides by 2003. The concentrated and successive treatment of ACCase and ALS inhibitors for control of barnyardgrass in rice led up to the resistance of E. oryzoides. Also, SU-herbicides like pyrazosulfuron-ethyl and imazosulfuron which effective to barnyardgrass can be bound up with the resistance of E. oryzoides. The ALS activity isolated from the resistant biotype of M. korsakowii to SU-herbicides tested was less sensitive than that of susceptible biotype. The concentration of herbicide required for 50% inhibition of ALS activity ($I_{50}$) of the SU-resistant M. korsakowii was 14-to 76-fold higher as compared to the susceptible biotype. No differences were observed in the rates of [$^{14}C$]bensulfuron uptake and translocation. Acetolactate synthase (ALS) genes from M. vaginalis resistant and susceptible biotypes against SU-herbicides revealed a single amino acid substitution of proline (CCT), at 197th position based on the M. korsakowii ALS sequence numbering, to serin (TCT) in conserved domain A of the gene. Carfentrazone-ethyl and pyrazolate were used mainly to control SU-resistant M. vaginalis by 2006 in Korea. However, the alternative herbicides such as benzobicyclone, to be possible to control simultaneously the several resistant weeds, have been developing and using broadly, because the several resistant weeds have been occurring simultaneously in the same fieled. The top ten herbicides by applied area in Korea have been occupied by products of 3-way mixture type including herbicides with alternative mode of action for the herbicide resistant weeds. Mefenacet, fentrazamide and cafenstrole had excellent controlling effects on the ACCase and ALS inhibitors resistant when they were applied within 2nd leaf stage.

Identification of Tumor Suppressor Loci on the Long Arm of Chromosome 5 in Primary Small Cell Lung Cancers (원발성 소세포폐암에서 염색체 5번의 장완에 위치한 종양억제유전자좌의 확인)

  • Cho, Eun-Song;Kim, Ho-Guen;Cho, Chul-Ho;Chang, Joon;Chung, Kyung-Young;Kim, Young-Sam;Park, Jae-Min;Kim, Sung-Kyu;Kim, Se-Kyu
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.1
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    • pp.49-59
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    • 2000
  • Backgrounds : Recent cytogenetic studies indicated that long of the long arm of chromosome 5 is a frequent event in small cell lung canær (SCLC), suggesting the presence of a tumor suppressor gene in its place. To map the precise tumor-suppressor loci on the chromosome arm for further positional cloning efforts, we tested 15 primary SCLCs. Methods : The DNAs extracted from paraffin-embedded tissue blocks with primary tumor and corresponding control tissue were investigated. Nineteen polymorphic microsatellite markers located in the long arm of chromosome 5 were used in the microsatellite analysis. Results : We found that ten (66.7%) of 15 tumors exhibited LOH in at least one of tested microsatellite markers. Two (13%) of 10 tumors exhibiting LOH lost a larger area in chromosome 5q. LOH was observed in five common deleted regions at 5q. Among those areas, LOH between 5q34-qter and 5q35.2-35.3 was most frequent (75%). LOH was also observed in more than 50% of the tumors at four other regions, between 5q14-15 and 5q23-31, 5q31.1, 5q31.3-33.3, and 5q34-35. Three of 15 tumors exhibited shifted bands in at least one of the tested microsatellite markers. Shifted bands occurred in 2.5% (7 of 285) of the loci tested. Conclusion : Our data demonstrated that at least five tumor-suppressor loci exist in the long arm of chromosome 5 and that they may play an important role in small cell lung cancer tumorigenesis.

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Developmental Genetic Analysis of Avian Primordial Germ Cells and the Application to Poultry Biotechnology

  • Kagami, H.
    • Korean Journal of Poultry Science
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    • v.28 no.2
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    • pp.135-142
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    • 2001
  • A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

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