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프탈레이트 에스터를 분해하는 Klebsiella sp. KBO222 의 Protocatechuate dioxygenase 유전자의 클로닝
진익렬,김상헌,김병오,서정훈,박영덕 경북대학교 유전공학연구소 1992 遺傳工學硏究所報 Vol.7 No.1
The recombinant DNA molecules of the insertion of the BamH1-digested fragment of Klebsiella chromosomal DNA into the E. coli plasmid vector molecule, pJL3-TB5, could be transformed into E. coli MC1061. The transformants appeared onto the selective medium with protocatechuate as a sole carbon source. Each colony was inoculated into this selective liquid medium, followed by a shaking overnight cultivation. The supernatant of one of the sonicated cultured cells did show the same activity of this protocatechuate dioxygenase with comparison to that of the original Klebsiella KBO222 cells. Further studies are being focused on the isolation of this gene from the transformant.
Ti-플라스미드 유전자 운반체에 의한 대두 형질전화 식물체에 있어서 β-Glucuronidase 유전자의 유전양상
김달웅,조은기,서득룡,오웬스, L . D . 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
To introduce β-glucuronidase (GUS) gene of Escherichia coli into plant cells using an Agrobacterium/binary vector system, we cocultivated shoot apices of soybean with Agrobacterium. After 3 weeks of culture on a culture medium containing 100 ㎎/L kanamycin, kanamycin resistant apices were selected. High GUS activities were detected in transformed explants by spectrophotometric assay. A tumefaciens ET₂[pGT100/EHA101] strain was more effective than the other strains for GUS expression from kanamycin resistant shoots. GUS was expressed in 43% of the cultured explants. It took only 6 weeks to regenerate the rooted plants. When transformants were self-pollinated, GUS gene was inherited to the next generation. Segregation ratio for GUS in T₂ suggests the insertion of a single copy of GUS.
한국인에서 사람 피필로마 바이러스 (HPV) 18 형 E6 유전자의 분리와 분석 및 대장균에서 18 형 E6 재조합단백질의 대량 생산
김현수,최인성,손우익,박순희,양은경,진승원 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
Human papillomavirus type 18 (HPV 18) has recently been closely linked with human cervical cancers. One of its early gene, E6 has been shown to code for a transforming protein. We analyzed the nucleotide sequences of three HPV 18E6 oncogenes isolated from korean women. One of them having exactly same nucleotide sequences to the reported transforming HPV 18E6 oncogene was cloned into pET-3a vector to construct a recombinant plasmid pET-18E6 to produce authentic size recombinant 18E6 protein in E. coli. We also isolated TrpE-18E6 fusion protein and generated antisera against it in rats. Using these antisera, we confirmed the expression of recombinant HPV 18E6 protein of authentic size from the recombinant plasmid pET-18E6 in E. coG. The expression level of recombinant 18E6 protein reached over 30% of total E. coli proteins but the protein was produced as inclusion bodies.
섬유질 Biomass 의 생물전환에 의한 대체에너지 생산에 관한 생물공학적 연구
이용현,하지홍,박중곤 경북대학교 유전공학연구소 1986 遺傳工學硏究所報 Vol.1 No.1
Anaerobic thermophilic bacteria Clostridium thermocellum can directly convert cellulosic biomass to ethanol with the simultaneous accumulation of fermentable sugars. To produce the alternative liquid fuel using above strain, the following works were carried out, (1) the cultivation technique of anaerobic thermophilic bacteria was established, (2) the cloning of cellulase gene was attempted, and (3) the cellulase complex was fractionated and their properties were determined.
Clostridium thermocellum 의 Cellulase 유전자의 Cloning
한성숙,이용현,하지홍,김욱한 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
A cellulase gene of Clostridium thermocellum was transferred to Escherichia coli by molecular cloning with pBR322. The gene was carried in n Hind III digested DNA sequence of about 1.8 kb. This Hind III fragment expressed activities on carboxymethyl cellulase (CMC) and on filter paper in E. coli. The expression of clostridial cellulose gene in E. coli was studied and compared with the products of cellulase genes in C. thermocellum.
대장균의 acetyl CoA carboxylase 유전자의 클로닝
송방호,홍순덕,박완 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
A defective lambda transducing phage carrying acetyl CoA carboxylase gene (fabE) from Escherichia coli chromosome (72 min on the current linkage map) has been isolated. A restriction map of the chromosomal region from defective transducing phage was established by digestion with combination of the restriction enzymes. No cleavage site for the enzyme EcoRI was found in this region. Restriction fragments were cloned from defective transducing phage into high copy number plasmid vector pACYC184 to generate hybrid plasmids which were capable of complementation of fabE temperature sensitive mutation. We show here that the fabE gene is located on a 3.4 megadalton BamHI-SalI fragment with a HindIII site, which lies within the 7.4 megadalton BglII fragment, by complementation analysis.
연구소보 발간 10 주년 연구논문 모음집:1985 년도 전문학술지 게재 논문;마찰반응계에서 전분질의 무증자당화에 관한 연구
이용현,조구형 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
The saccharification of uncooked starch was carried out in an attrition-coupled reaction system. The mechanical movement of the milling-attrition media contained in the starch slurry enhanced the hydrolytic action of the starch Iiquifying and saccharifying enzymes significantly. I he enhancement may be due to the attrition effect rather than the milling effect. It was found that [tie enzymes were relatively stable even under the attrition state. In the case of the naked barley powder, both saccharification rate and sugar yield were improved compare with conventional cooking saccharification method. The effectiveness of the various milling media, such as, glass bead, polyacetal bead, and teflon bead were evaluated. Even at the concentration as high as 38 % (w/v). the starch can be effectively hydrolyzed in an attrition coupled reaction system. Although further studies need to be conducted to evaluate [lie feasibility of its commercialization, this novel system provides an improved method for the saccharification of starch without cooking.
Agrobacterium tumefaciens 를 이용한 Petunia hybrida 엽조직내로의 GUS (β-Glucronidase) 유전자의 도입 가능성
정재동,지선옥,김창길,권명용 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
Petuina kybrida was transformed with GUS gene by cocultivation of leaf disc with Agrobacterium tumefacines. Petunia leaf showed resistant response to the concentration of 100 ㎎/L kanamycin. From the petunia leaf dises(about 5×5 ㎜) cocultivated with Agrobacterium suspension cultures for 48 hours in MS medium containing 1.0 ㎎/L 2,4-D and 0.2 ㎎/L BAP, kanamycin resistant callus were more effectively induce. Shoots regenerated from the callus when cultured on MS medium containing 1.0 ㎎/L BAP, 0.1 ㎎/L NAA, 200 ㎎/L kanmycin, 500 ㎎/L carbenicillin, 30 g/L sucrose and 8 g/L agar. 59% of regenerants showed positive color response when reacted with X-glu.