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Blue/White screening이 가능한 새로운 T-vector pBX223계의 개발 및 응용
이동우,이도부,이혜민,한상훈,윤병수 경기대학교 사회과학연구소 2005 경기대학교 기초과학논문집 Vol.18 No.1
To facilitate the cloning of PCR products, T-vector systems have been developed and widely used. However, PCR product cloning with established T-vector systems often produces colonies with a self-ligated or uncut vector, leading a wrong selection of clone in Blue/White screening which distinguishes clones with target insert DNA by the color of clonies. Therefor, additional screening tests are required to confirm PCR product insertion. To make up for the defect of established T-vector systems, we developed pBX223 system composed of three T-vector, pBX223W1, pBX223W2, and pBX223B. There vector enable not only Blue/White screening but also anticipating the color of the positive clones when the sizes of PCR products are known. In actual tests, the accuracy rate of pBX223W1, pBX223W2, pBX223B were 87%, 87%, and 86%, respectively. In conclusion, pBX223 T-vector system may be useful to the cloning PCR products and provide an additional advantage in the screening process if positive clones.
대장균에서 Chlamydia psittaci MOMP 유전자의 과발현과 순수분리
하정순,이도부,한상훈,임윤규,윤병수,Ha, Jung-Soon,Lee, Do-Bu,Han, Sang-Hoon,Lim, Yoon-Kyu,Yoon, Byoung-Su 대한수의학회 2006 大韓獸醫學會誌 Vol.46 No.1
Generally known psittacosis or ornithosis is a disease of birds caused by the bacterium Chlamydia psittaci. Humans are accidential hosts and are most commonly infected from avian sources. It raises hepatitis or neurosis. As major outer membrane protein (MOMP) of Chlamydia psittaci has been known to play a role in the avoidance of host immune defenses, research on developing a Chlamydia vaccine has focused on the MOMP. In this study, the gene encoding the major outer membrane protein (MOMP) of the Chlamydia psittaci strain 6BC was cloned and expressed in Escherichia coli strain M-15. The recombinant DNA was cloned by fusion prokaryotic expression vector pQE30-GFPII. Expression of the recombinant protein was performed in E. coli and was induced by IPTG. The size of expressed recombinant protein is 74.220 kDa (MOMP, 43.260 kDa; GFP expression region, 30 kDa; $6{\times}His$ tag, 960Da). This protein was purified by using his-tagging-inclusion body. Recombinant protein was reconfirmed through ELISA test and western blot with antibody against pQE30-GFPII. It will be useful antibody development.
버들치(Rhynchocypris oxycephalus)의 vitellogenin 생성량 측정을 이용한 수중 estrogen휴 화합물의 정량분석법에 관한 연구
김을환,이도부,한상훈,윤병수 경기대학교 부설 기초과학연구소 2005 경기대학교 기초과학논문집 Vol.18 No.1
Vitellogenin(Vtg), a phospholipoglycoprotein precursor of egg yolk, is synthesized and secreted from the liver in response to estrogens in female fish. Vtg is normally undetectable in the blood of male fish, but can be induced by exposure to chemicals possessing estrogenic activity. Thus, the presence of Vtg in blood of male fish can serve as a useful biomarker for assessing previous exposure to estrogenic compounds. In the present study, Vtg was abnormally expressed in Rhynchocypris oxycephalus treatment of Estradiol Benzoate (E_(2)). As the result, it was found that the level of Vtg in blood from R. oxycephalus was increased by treated quantity of E_(2) with dose-effect manner.