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동일한 시료에 대한 국내 기관간의 STR 분석결과 비교 : STR 유전자좌 분석법의 표준화 설정을 위하여
박종태,신경진,양윤석,우광만,이숭덕,이승환,이정빈,정연보,조승희,한길로,한면수,홍승범 大韓法醫學會 2001 대한법의학회지 Vol.25 No.1
This paper described a collaborative exercise intended to see what kinds of short tandem repeat (STR) loci are used in different DNA typing laboratories in Korea and to compare their results for the demonstration whether uniformity of DNA profiling results from different laboratory could be achieved in Korea Laboratories were asked to test five tissue DNAs using methods routinely used in each laboratory and to report the results to the coordinating laboratory. The exercise demonstrated that each laboratory was using different STR loci for the typing with different STR numbers,2 VNTRS,36 STRs and amelogenin in total, and the direct comparison of the results from all the laboratory for the 18 loci could not be done as only one laboratory submitted typing results. Among 21 loci for which several laboratories submitted typing results, results for 14 loci were the same and results for the other 7 loci were different depending on the participating laboratory. D1S80, F13A01, D16S539, D21S11, D18S51, D3S1744 were the loci with different typing results. Even in the cases where commercial kits were used, the results were not the same depending on the machines used, that is the capillary electrophoresis or the gel based electrophoresis. The reason for the different results, points about the standardization of the methods arid the profiling data were described.
동종 골수 이식한 백혈병에서 단반복 유전자로 살펴본 혼합 키메라 현상의 의의
장대영,이정신,서철원,이규형,이제환,지현숙,박찬정,한면수,최동원,김정균,최성준,김성배,김상위,김우건,김상희 대한조혈모세포이식학회 1998 대한조혈모세포이식학회지 Vol.3 No.1
연구배경 : 동종 골수 이식한 백혈병에서 키메라 현상의 분석은 착상, 질병 재발과 이식 거부를 이해하는데 도움을 줄 수 있다. 최근 STR을 PCR로 증폭한 검사법에 의해 매우 민감하고 유용하게 혼합 키메라 현상을 검출할 수 있다고 보고되고 있으나, 이 방법의 민감도와 임상적 의의는 아직 완전히 정립되지는 않았다. 따라서 PCR-STR검사법이 혼합 키메라 현상을 민감하게 검출하는지 보고, 동종 골수 이식 후 혼합 키메라 현상의 변화 양상을 관찰하고, 키메라 현상의 임상적 의의를 보고자 하였다. 방법 : 형제로부터 동종 골수 이식한 급성 백혈병 6명과 만성 백혈병 2명의 모두 8명을 대상으로 하였다. 44개 골수 검체에서 phenol/chloroform방법에 의하여 단핵세포 DNA를 추출하였다. DNA는 CTT 삼중체, vWA와 amelogenin시발체를 이용하여 증폭하였다. 증폭 산물은 5% 폴리아크랄아미드 젤에서 전기영동 뒤 은 염색으로 확인하였다. 2명의 DNA를 여러 비율로 혼합하여 상기 검사를 하여 검출 가능 최소 농도를 결정하였다. 결과 : 상기 검사법의 검출 가능 최소농도는 0.25%였다. 혼합 키메라 현상은 8명중 4명에서 관찰되었다. 혼합 키메라 현상은 보인 1명에서 급성 및 만성 이식편대 숙주 질환이 나타났고, 완전 공여자 키메라 현상을 보인 1명에서 만성 이식편 대 숙주 질환이 나타났다. 점진적으로 증가하는 혼합 키메라 현상을 보인 2명은 이식 후 12개월에 질병 재발을 보였으나, 완전 공여자 키메라 현상을 보인 경우엔 재발이 관찰되지 않았다. 결론 : PCR-STR검사법은 동종 골수 이식 후 남아 있는 환자 세포를 검출하고 키메라 현상을 관찰하는데 유용한 방법이다. 또한 이 검사법은 동종 골수 이식 후 재발의 조기 진단, 이식편 대 숙주 질환의 평가와 면역 관용에 임상적으로 응용할 수 있으리라 생각한다. Background : Chimerism analysis after allogeneic bone marrow transplantation (BMT) for leukemia could be helpful understanding of the early marrow engraftment, disease relapse, and graft rejection. Recently, a PCR technique which amplifies short tandem repeats(STR) has been reported to be highly sensitive and reliable in detecting mixed chimerism. But its sensitivity and clinical significance has not been established. The purpose of this study was firstly, to confirm whether a PCR-STR in highly sensitive enough ti assess mixed chimerism, secondly to monitor the changing patterns of mixed chimerism after allogeneic BMT, and lastly to determine correlation between the chimeric status and the clinical outcome. Methods : A study was made eight patients (six with acute leukemia and two with chronic leukemia) who underwent unmanipulated allogeneic BMT along with matched donors. DNA was extracted from mononuclear cells of 44 bone marrow samples using a phenol/chloroform method. Amplification of DNA was done using CSFIPO-TPOX-TH01(CTT) triplex with or without vWA or amelogenin primer. The amplified product was separated on 5% polyacrylamide gel and was confirmed by silver staining. The sensitivity of CTT triplex method was determined by mixing DNA from two person in serial proportion using the same method. Results : The sensitivity for CTT triplex method was 0.25% Mixed chimerism was documented in 4 of 8 patients. ONe of four patients with mixed chimerism developed acute and chronic graft versus host disease (GVHD) and one of four patients with complete donor chimerism also developed chronic GVHD. Two patient with progressive mixed chimerism relapsed at 12 months post-BMT, whereas no patient with complete donor chimerism has relapsed. Conclusion : The study has found that PCR-STR was an effective method for the detecting residual host cell and monitoring the chimeric status after allogenic BMT. PCR-STR can also be clinically applicable in early prediction of relapse, appropriate assessment of GVHD, as well as tolerance after allogeneic BMT.
Continuous Ethanol Production Using Immobilized Baker's Yeast
Han, Myun Soo,Ha, Sang Do,Chung, Dong Hyo 한국산업미생물학회 1991 한국미생물·생명공학회지 Vol.19 No.4
효모를 Na-alginate에 고정화한 후 연속반응기를 이용한 glucose 빌효로 에탄올을 생산하였다. 그 결과 고정화 효모의 활성화 시간은 20∼25시간이었다. 연속발효에서 고정화효모의 온도안정성은 30∼37℃였으며 pH 안정성은 pH 4.0∼pH 8.0, 최적희석속도는 0.2h^-1이었고 에탄올생산 최적 당농도는 15%였다. 최적조건에서 에탄올수율은 0.23, 생산된 에탄올 농도는 33.90g/l 그리고 에탄올생산성은 7.12g/l·h로 각각 나타났다. Ethanol production by calcium alginate-immobilized baker's yeast was studied in the continuous shaked-flask reactor (CSFR) using glucose medium as a feed. Immobilized cells were stable at 30∼37℃ and pH 4∼8. Fermentation characteristics of immobilized baker's yeast were examined changing the initial glucose concentration employed were 50, 100 and 150 g/l, respectively. It was investigated that the influent glucose concentration and the dilution rate have an influence on the ethanol fermentation characteristics at steady state in continuous culture of immobilized baker's yeast. The optimum conditions for high ethanol productivity and low residual glucose output in ethanol prodution were shown to be 0.2h^-1 for the dilution rate and 150g/l for the influent glucose concentration. The maximum ethanol productivity, ethanol yield, specific growth rate and glucose conversion rate were around 7.12g/l·h, 0.23, 0.366g/l·h and 78.43, respectively.
Han, Myun Soo,Hong, Seung Beom,Choi, Sang Kyu,Cho, Youl Hey,Jin, Han Jun,Kwak, Kyoung Don,Kim, Wook,Kim, Jong Bong,Son, Su Min 한국유전학회 2002 Genes & Genomics Vol.24 No.1
We analyzed variations at thirteen Combined DNA Index System (CODIS) short tandem repeat (STR) loci (CSF1PO, FGA, THO1, TPOX, vWA, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51, and D21S11) in a sample from 130 unrelated individuals in the Korean population. Allele and genotype frequencies were determined using commercial PCR amplification kits. The Exact Test demonstrated that all loci were found to be no deviations from Hardy-Weinberg expectations (P>0.05). The distribution pattern of allele frequencies of the 13 STR loci reflected a common genetic affinity in contemporary populations from east Asians, but the Koreans are more closely related with the Chinese than to the Japanese and Vietnamese in east Asians surveyed so far. Based on the results of allele frequencies of the 13 CODIS STR loci, Koreans studied here were the most significantly different from those of African-American and Caucasian. For forensic testing, the power of discrimination (PD) index ranged from 0.760 at TPOX to 0.961 at FGA. The combined probability of match (PM) calculated from all 13 core CODIS STR loci was 1.94×10 exp (-14), which is highly informative. Therefore, the Korean 13 CODIS STR data could be useful for the regional specific and prerequisite references to the forensic community.
Studies on the Immobilization of Saccharomyces cerevisiae for Ethanol Production
Han, Myun Soo,Ha, Sang Do,Chung, Dong Hyo 한국산업미생물학회 1991 한국미생물·생명공학회지 Vol.19 No.4
효모를 Ca-alginate에 고정화하여 회분발효에서 glucose로부터 에탄올을 생산하여 다음의 결과를 얻었다. 100g wet weight/l(4.3×10 exp(9) cell/l)의 효모를 pH 7.0, 2% 농도의 Ca-alginate에 고정화하였다. 10% beads volume이 에탄올 생산에 최적이었고 30일(720시간) 동안 bead의 수명이 지속되었다. 회분식 발효에서 온도안정성은 고정화 효모의 경우 30∼40℃였으며 free cell의 경우 30∼37℃였다. pH 안정성은 pH 4.0∼9.0였으며, 에탄올생산 최적 당농도는 15%였다. 최적조건에서 에탄올수율은 0.45, 생산된 에탄올 농도는 67.5g/l 그리고 에탄올 생산성은 1.99g/l·h로 각각 나타났다. Ethanol production by calcium alginate-immobilized baker's yeast (Saccharomyces cerevisiae) was studied in the batch fermentation using glucose medium as a feed. Immobilied cells were stable between 30℃ and 40℃ whereas free cells were stable between 30℃ and 37℃. The beads were showed constant ethanol productivity during 720 hours (30 days) over. Fermentation characteristics of immobilized baker's yeast were examined changing the initial glucose concentration of broth in fermentation. Initial glucose concentrations employed were 50, 100, 150 and 200 g/l, respectively. In 15% gucose medium, maximum specific growth rate, maximum ethanol yield and ethanol concentration were observed as 0.092h^-1, 0.45, 67.5g/l, respectively.