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Cho, Yong K.,Choi, Young K. Institute of Genetic Engineering Changwon National 1998 Gene and Protein Vol.2 No.1
The data show that combining ion-exchange with reversed-phase partition chromatography accomplished purification of NADH that neither technique alone could provide. When the product NADH was stored in a dry vehicle, it was found to give a single, major peak when analyzed by reversed-phase analytical HPLC.
미나리 체세포배에서 특이적으로 발현되는 EF1-αc DNA의 분리와 특성파악
빈철구 Institute of Genetic Engineering Changwon National 1997 Gene and Protein Vol.1 No.1
This experiment was carried out to isolate and characterize the elongation factor cDNA expressing during somatic embryogenesis in water dropwort (Oenanthe stolenifera DC.) An optimal concentration of 2,4-D was 1mg/l for induction of embryogenic callous (EC) from meristematic tissue of water dropwort. Somatic embryos were relatively well induced from EC in hormone free medium. The cDNA of elongation factor, EF₁-α, which was expressed during somatic embryogenesis, was isolated by PCR (polymerase chain reaction) using a primer which was synthesized based upon the DNA sequence of tomato EF₁-α. The cDNA of EF₁-α was partially sequenced by direct sequencing method of PCR product. The sequence of the EF₁-α was highly homologous with that of carrot (ca. 85% homology) , tomato (ca. 80%), and Arabidopsis (ca. 80%). Tissue specific expression of the EF₁- α DNA was tested by the RNA/PCR. It was confirmed that EF₁-α DNA was actively expressed in the somatic embryo and the embryogenic callus, but not in the nonembryogenic callus. Genormic Southem analysis was carried out using the EF₁-α DNA as a probe, and several DNA bands were observed. It was suggested that the EF₁-α DNA was a multigene family.
Optimization of Callus Formation from Stem,Leaf and Root Explants of Tomato(Lycopersicon esculentum)
Cho,Un Haing Institute of Genetic Engineering Changwon National 1997 Gene and Protein Vol.1 No.1
Effects of various plant hormones were examined for optimal callus induction from various tomato tissues. The best results were obtained on MS medium supplemented with 0.001-l0μM 2,4-D, 1.0-10.0μM insole-3-acetic acid(IAA) and 10.0μM zeatin from stem explants, with 1.0-10.0μM 2,4-D from leaf explants, and with 1.0μM phenylacetic acid(PAA ) and 10.0μM zeatin from root explants. The protocol Is suitable for preservation of gemplasm of useful traits selection of variants.
α-Functionalized Benzylamine의 합성
金興載,曺壽東,安撤鎭,朱尤洪,申東守 Institute of Genetic Engineering Changwon National 2000 Gene and Protein Vol.4 No.-
α-Functionalized benzylamine(Nu=OAcm OMe, N_3)은 유기합성의 출발물질 또는 synthon으로서 매우 유용할 것이다. 본 연구에서는 benzylphthalimide를 137℃, chlorobenzene 용매 하에서 NBS/NaOAc/HOA조건을 사용하여, 높을 수율로 phthalimide-benzyl acetate을 얻는 방법을 개발하였다. 마지막으로, phthalimide-benzyl acetate을 NH_2NH_2/HOAc 조건 하에서, amino(phenyl)methyl acetate ⑴을 합성하였다. α-Functionalized benzylamine(Nu=OAc, OMe, N_3)will be very useful as a starting material or synton in organic synthesis. We have investigated the first synthetic methodology of phthalimide-benzyl acetate form benzylphthalimide, which are prepared in NBS/NaOAc/HOAc in chlorobenzene at 137℃ for 12 hr in good yield. Finally, amino(phenyl)methyl acetate ⑴ was synthesized from phthalimide-benzyl acetate under NH_2NH_2/HOAc.
Pseudomonas sp. BCNU 171균에 의한 유기용매 내성 단백질의 탐색
이승범,정미연,김미림,박정욱,김민정,주우홍 Institute of Genetic Engineering Changwon National 1999 Gene and Protein Vol.3 No.1
Pseudomonas sp. was screened from wastewater. The strain was able to grow in the presence of 10% (v/v) toluene. The toluene resistance was significantly improved by ?? ion. Tricine-SDS-PAGE showed some changes in the protein profile of the strain when the toluene stress was given. Notably, an organic solvent tolerant protein, OSP-7 was detected by the toluene stress.
재조합 효모로부터 생산된 glucose oxidase의 정제
이대희,이승철,황용일 Institute of Genetic Engineering Changwon National 1999 Gene and Protein Vol.3 No.1
The yeast expression vector, pGAL-GO02, that consist of a GAL10 promoter, α-amylase signal sequence, mature glucose oxidase(GOD) coding sequence and glucose oxidase terminator, was used for production of glucose oxidase from Aspergillus niger. The overproduction and high level secretion of GOD from A. niger in Saccharomyces cerevisiae 2805 was carried out by cloning GOD gene. Transformed S. cerevisiae with pGAL-GO02 produced 8.7unit/ml of GOD that is c.a. 5 times higher than 1.7unit/ml in A. niger on YPDG media. For isolation and purification of secreted GOD, culture broth was microfiltrated. The microfiltrated GOD was further purified by dialysis and ion-exchange chromatography. The final product is more than 95% pure and the specific activity was 8.6unit/mg that is increased about 4.7-fold than 1.83unit/mg in crude extract. Molecular weight of the purified GOD was c.a. 122 kDa by hyperglycosylation in yeast and molecular weight of treated GOD with Endoglycosidase H is 69 kDa.
대황 및 황련에탄올 추출물 처리에 의한 시설채소산물의 품질 변화
박우포 Institute of Genetic Engineering Changwon National 1999 Gene and Protein Vol.3 No.1
To maintain the quality characteristics of vegetables, cucumber, zucchini, green pepper and strawberry were dipped in Korean medical herb extracts (KMHE) such as Rheum palmatum L. or Coptis chinensis Franch with different concentrations. Dipping in 1000 ppm KMHE solution was suitable for the reduction of microbial load on cucumber, and 500 ppm was appropriate for others. Dipping time was adequate for produce about 10 minutes, and the temperature of dipping solution had a limited effect on the microbial load reduction. Antimicrobial activity of KMHE was maintained for cucumber and zucchini packaged with 30 μm low density polyethylene film during storage at 10℃.
Toluene내성세균 Pseudomonas sp. BCNU 154에 의한 BTEX의 분해
성은미,이호원,주우홍 Institute of Genetic Engineering Changwon National 1999 Gene and Protein Vol.3 No.1
The biodegradative potentialities of a toluene-tolerant Pseudomonas sp. BCNU 154, isolated from wastewater, were investigated. Among 16 aromatic substrates tested, cumene, cyclohexane, ethybenzene, p-xylene, m-xylene, toluene and diphenylether were metabolized. Pseudomonas sp. BCNU 154 degrades aerobically toluence, ethybenzene, p-xylence and cumene. With toluene, complete degradation occurred after 12 hours but with p-xylene and cumene, and with ethybenzene, 90 and 75% degradation occurred after 12 hours of incubation, respectively.
Cho, Un-Haing Institute of Genetic Engineering Changwon National 1998 Gene and Protein Vol.2 No.1
토마토 조직배양에 있어서 ethylene 형성억제제 (AOA 및 TIBA) 및 활성억제제 (AgNO₃)와 조직의 노화억제제 (Vitamin C, Vitamin E 및 putrescine)들이 callus 및 shoot 형성에 미치는 영향에 대해 조사하였다. 배지에 ethlene 억제제와 노화억제제의 첨가는 배양된 잎절편으로부터 callus형성을 억제하였지만 shoot의 형성을 촉진하였으며 유도된 callus를 Vitamin C가 첨가된 배지에 계대배양했을 때에도 shoot 형성을 증가 시켰다. 본 실험결과들은 배양된 조직에 있어서 callus 형성을 위해서는 ethylate 형성이나 조직의 노화과정이 필요하지만 기관분화발달시기에 있어서는 ethylene 형성의 억제와 조직의 노화억제가 필요함을 보여준다.
Kim,Jin Kyoo,Lee,Soo O,Kim,Chul Hong Institute of Genetic Engineering Changwon National 1997 Gene and Protein Vol.1 No.1
Recombinant Fc-hinge fragment of murine immunoglobulin G1(IgG1) was expressed in Escherichia coli and intracellular sites of expressed protein were determined for this purpose, three different fractions were derived from periplasmic space, cytoplasm and inclusion body respectively in E. coli and then were analyzed using SDS-PACE, immunoblotting and staphylococcal protein A affinity chromatography. The result show that recombinant Fc-hinge fragment were preferentially expressed in cytoplasm. In addition, purified Fc-hinge fragment retained staphylococcal protein A binding activity, suggesting its biological activity may be as same as native murine IgG1.