• Title/Summary/Keyword: 유전적 재조합

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Solid-Phase Refolding Technology in Recombinant Proteins Recovery: Application Examples to Various Biopharmaceutical Proteins (유전자재조합 단백질 회수 공정에서의 고체상 재접힘 기술: 여러 바이오의약 단백질에의 적용 사례)

  • Kim, Min Young;Suh, Chang Woo;Kim, Chang Sung;Jo, Tae Hoon;Park, Sang Joong;Choi, Won Chan;Lee, Eun Kyu
    • Korean Chemical Engineering Research
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    • v.43 no.2
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    • pp.187-201
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    • 2005
  • Bioprocessing technologies utilizing 'biorecognition' between a solid matrix and a protein is being widely experimented as a means to replacing the conventional, solution-based technology. Frequently the matrices are chromatographic resins with specific functional groups exposed outside. Since the reactions of and interactions with the proteins occur as they are attached to the solid matrix, this 'solid-phase' processing has distinct advantages over the solution-phase technology. Solid-phase refolding of inclusion body proteins uses ion exchange resins to adsorb denaturant-dissolved inclusion body. As the denaturant is slowly removed from the micromoiety around the protein, it is refolded into a native, three-dimensional structure. Once the refolding is complete, the folded protein can be eluted by a conventional elution technique such as the salt-gradient. This concept was successfully extended to 'EBA (expanded bed adsorption)-mediated refolding,' in which the denaturant-dissolved inclusion body in whole cell homogenate is adsorbed to a Streamline resin while cell debris and other impurity proteins are removed by the EBA action. The adsorbed protein follows the same refolding steps. This solid-phase refolding process shows the potential to improve the refolding yield, reduce the number of processing steps and the processing volume and time, and thus improve the overall process economics significantly. In this paper, the experimental results of the solid-phase refolding technology applied to several biopharmaceutical proteins of various types are presented.

Genetic Recombination of Brevibacterium lactofermentum by Protoplast Fusion (Brevibacterium lactofermentum의 원형질체 융합에 의한 유전자 재조합)

  • 이혜경;최순영;윤윤경;이영하;민경희
    • Korean Journal of Microbiology
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    • v.28 no.2
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    • pp.98-103
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    • 1990
  • Brevibacterium lactofermentum SWA (arg trp) and B. lactofermentum SWB (met ser) were obtained from UV and NTG treatment. The rates of protoplast formation by B. lactofermentum SWA and SWB were 99.93% and 99.98%, respectively when each strain was treated with penicillin G in mid exponential growth phase, followed by incubation with 400 $\mu\textrm{g}$/ml of lysozyme in lysis fluid supplemented with 0.4M sucrose. Frequencies of protoplast regeneration in B. lactofermentum SWA and B. lactofermentum SWB were 9.27% and 10.32% respectively, on regeneration medium containing 0.5M sodium succinate, 50 mM $Mg^{2+}$, and 3% PVP. In intraspecific protoplast fusion between B. lactofermentum SWA and B. lactofermentum SWB, fusion frequency of $2.30\times 10^{-5}$ was observed by using the 100mM $CaCl_{2}$ and 30% PEG 6,000 in fusion fluid. Relative recombinant frequencies in each marker by means of selective media could be used for genetic analysis.

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Comparison of Regeneration Conditions in Seven Pepper (Capsicum annuum L.) Varieties (7종의 고추(Capsicum annuum L.) 재분화 조건 비교)

  • Min-Su Kim;Yun-Jeong Han;Sharanya Tripathi;Jinwoo Kwak;Jin-Kyung Kwon;Byoung-Cheorl Kang;Jeong-Il Kim
    • Korean Journal of Plant Resources
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    • v.36 no.5
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    • pp.527-539
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    • 2023
  • Pepper (Capsicum annuum L.) is an important vegetable and spice crop that has been cultivated worldwide. Pepper fruits have unique taste and aroma, providing a variety of antioxidants and compounds important for human health, which makes a high economic value. In addition, there is a high demand for new pepper varieties, according to consumer's preference. However, pepper is a recalcitrant plant for in vitro tissue and organ differentiation and plant regeneration, which makes it difficult to develop demanded varieties using newly developed technologies such as genetic engineering and gene editing. In this study, tissue culture and regeneration conditions were investigated using seven pepper varieties that were obtained from the core-collection of Seoul National University. We observed callus and bud induction and shoot formation using several media composition composed of different cytokinins and auxin concentrations. As a result, it was found that there were differences in callus induction and shoot formation of each variety depending on the hormone composition, and the highest regeneration was shown when the medium containing Zeatin Riboside and the petioles of seedlings were used. In particular, out of seven pepper varieties, CMV980 exhibited a higher regeneration efficiency (approximately 48%) than other varieties, followed by Yuwolcho. Therefore, this study provides CMV980 and Yuwolcho as good candidates that can be used for pepper transformation, which might contribute to the development of various varieties through gene editing technology in the future.

생명공학과 담배 및 다른 작물들의 응용 현황

  • 박성원
    • Proceedings of the Korean Society of Tobacco Science Conference
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    • 2001.05a
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    • pp.72-78
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    • 2001
  • 농업 유전공학 기술은 생산성 향상, 환경보전, 식품의 안정성 및 품질향상에 기여함은 물론 농업의 경쟁력을 높일 수 있는 유일한 대안으로 인식되고 있다. 전 세계적으로 유전자 재조합 작물의 경작지는 2000년 한해동안 지난해 같은 기간에 비해 11% 증가하였으며 이는 1996년 대비 25배 증가하였고, 선진국과 개발도상국은 각각 2%, 51% 1999년 대비 증가하였다. 1983년 유전자 재조합에 의한 식물의 형질전환이 성공한 뒤 종묘업계는 형질전환 종자개발과 보급에 열중하고 있으며 종자시장에 형질전화 품종이 차지하는 비율은 2000년 30억 달러에서 2010년이면 전체의 60%인 200억 달러에 이를 것으로 전망된다. 1995년 제초제 저항성 콩(라운드업레디콩)이 농가에 보급된 이후 2000년 형질전환품종 재배면적이 3990만 ha에 이르렀고 1997년 미국과 캐나다는 옥수수, 대두, 면화, 감자, 유채 등의 형질전환 품종 재배로 각각 3억1400만 달러, 5300만 달러를 벌어들였음. 형질전환 품종의 보급 증가속도는 소비자들의 GMO에 대한 거부반응으로 다소 주춤한 상태이다. 그러나 최근 종자회사들은 생태계 위해성 논란을 피해갈 수 있는 연구로 이러한 상황을 돌파하려 하고 있다. 우리나라에서도 유전자변형 생물체에 관한 법률이 제정되었으며 많은 대학과 연구소에서 형질전환 연구가 꾸준히 이루어지고 있고 최근 제초제 저항성 벼와 바이러스 저항성 감자가 개발돼 GMO 안정성 점검에 들어가 있고, 살충성 배추, 혈압강하 토마토, 지방산 강화 들깨, 병저항성 고추 등도 실험실과 포장에서 재배되고 있다. 이르면 4-5년 뒤 형질전환 작물들이 농가에 보급될 전망이다. 이처럼 체크 툴은 Firewall의 수비능력을 보강하는 위치에 있다고 생각할 수 있다.다. 4 장에서는 3장에서 제기한 각각의 문제점에 대해 RAD 의 관점에 비추어 e-business 시스템의 단기개발을 실현하기 위한 고려사항이나 조건 해결책을 제안한다. 본 논문이 지금부터 e-business 를 시작하려고 하는 분, e-business 시스템의 개발을 시작하려고 하는 분께 단기간의 e-business 실현을 위한 하나의 지침이 된다면 다행이겠다.formable template is used to optimize the matching. Then, clustering the similar shapes by the distance between each centroid, papaya can be completely detected from the background.uage ("Association of research for algorithm of calculating machine (1992)"). As a result, conventional NN and CNN were available for interpolation of sampling data. Moreover, when nonlinear intensity is not so large under the field condition of small slope, interpolation performance of CNN was a little not so better than NN. However, when nonlinear intensity is large under the field condition of

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Evaluation on the Lost Prestressing Force of an External Tendon Using the Combination of FEM and HGA: II. Experimental Verification and Field Applications (FEM과 HGA의 조합을 이용한 외부 긴장재의 손실 긴장력 평가: II. 실험적 검증 및 현장적용)

  • Jang, Hang-Teak;Noh, Myung-Hyun;Park, Kyu-Sik;Park, Taehyo
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.13 no.5 s.57
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    • pp.121-132
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    • 2009
  • This paper introduces an experimental verification and a field application of the proposed technique using the combination of FEM and HGA about the loss prestressing force of an exteranl tendon by above same authors. The vibration tests have been conducted by using a laboratory models and the externally prestressed tendon at the field and the natural frequencies are extracted from the vibration tests. The proposed technique based on the extracted natural frequencies is applied. It is seen that the errors in the tension and lost prestressing force by proposed technique are about 4% from a laboratory model test. For the model verification at field, exact modeling has beem made with Rayleigh damping. It is seen that the error in the tension by proposed technique is less than 1% and the estimated lost prestressing force converges less than the exact value.

Restriction Mapping of Cloned Pullulanase Gene and Property of Pullulanase Produced in Escherichia coli (pYKL451) and Klebsiella pneumoniae NFB-320 (Klebsiella pneumoniae NFB-320의 Pullulanase 유전자의 제한효소 분석과 효소학적 특성)

  • Yu, Ju-Hyun;Chung, Kun-Sub;Kong, In-Su;Lee, Jung-Kee
    • Microbiology and Biotechnology Letters
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    • v.15 no.6
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    • pp.436-440
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    • 1987
  • Pullulanase gene (pul) of Klebsiella pneumoniae NFB-320 which was cloned previously in Escherichia coli with plasmid pBR322. The gene was analyzed with various restriction enzymes. The cloned gene was contained within n 10 kb BamHI DNA fragment. We constructed the restriction map of the hybrid plasmid pYKL451. The optimum temperatures for pullulanases produced in E. coli (pYKL451) and K. pneumoniae NFB-320 were almost the same, 50-55 $^{\circ}C$. The optimum pHs for the reaction of the enzymes produced by E. coli (pYKL451) and K. pneumoniae NFB-320 was 6.0. Both enzyme preparations were stable under the range of pH 5.0 to 10.0 when those were kept at 40 $^{\circ}C$ for 90 min and were stable until 40 $^{\circ}C$ when allowed to stand for 1hr at various temperatures.

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Comparative Study of DNA Fragment According to Steps of Genetically Modified Soybean Processed Food (대두 가공식품의 공정단계별 유전자재조합체(GMO) 단편의 검출확인 및 비교 연구)

  • Hwang, Sun-Wook;Lee, Cheol-Su;Nam, Young-Suk;Kim, Su-Bok;Oh, Duk-Hwan;Kim, Young-Chan
    • Journal of Food Hygiene and Safety
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    • v.18 no.4
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    • pp.211-217
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    • 2003
  • To discriminate between the genetically modified soybean processed foods which were Korean traditional foods as Beansprout, doenjang, Beancurd (Tofu) and the unmodified one. we had analyzed comparatively that the loss degree of inner DNA about denaturalization factors in process step as heat or pressure and decision of suitable PCR primer by size. As a result of having compared about ${\beta}$-actin was 160bp, 335 promotor was 130 bp and NOS terminator was 132 bp effective. As a result of having checked a loss degree of a gene, as for the bean curd, DNA was mostly preserved well, and the loss of DNA along the processing process was hardly observed by a processing process. Most DNA of beansprout have moved to trunk after germination stage, and the appropriate analysis part was judged as the trunk. And the doenjang showed a detection difference of DNA by an operation of an enzye among self-life periods. Besides, after 50days, insertion gene was destoryed entrely so that detection was not possible.

Establishment of a New Breeding Scheme for Rapid Release of Variety Using bar Gene Transformed Rice (형질전환 벼 이용 품종 조기개발 체계 확립)

  • Won, Yong-Jae;Yi, Gi-Hwan;Cho, Jun-Hyeon;Ko, Jong-Min;Park, Hyang-Mi;Han, Chang-Deok;Yang, Sae-Jun;Kim, Soon-Chul;Nam, Min-Hee
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.7-11
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    • 2004
  • This study was carried out to establish a new breeding scheme which is connected with conventional breeding method and anther culture method. To develop a herbicide resistant and direct seeding rice, $F_1$ plants were subjected to anther culture and regenerated plants from 5 crosses were studied to confirm the introduction of bar gene. After PCR analysis, we selected 227 plants which were carrying herbicide resistance gene (bar) out of 1,508 regenerated plants from anther culture. Among 169 $A_2$ lines carrying herbicide resistant gene from 5 crosses including YR23235 (Dongjin Ds3(Ba $r^{R}$)/ Milyang165), 42 lines that had superior agronomic characters were selected for further research. Among them, YR23235Acp79 which showed herbicide resistance, direct seeding adaptability and superiority in major agronomic characters was named Milyang 204. This breeding scheme proved that the anther culture of $F_1$ plants crossed between transformant and cultivar or transform ant alone could be utilized in breeding programs for a rapid progeny fixation and development of a variety.y.

Protein Therapeutics Industry Analysis and its Business Opportunities (단백질 의약품의 사업화기회분석)

  • Choi, Yun-Jeong
    • Proceedings of the Korea Contents Association Conference
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    • 2007.11a
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    • pp.478-481
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    • 2007
  • To date, modern medicine has relied heavily on synthetically or chemically produced drugs to treat or prevent diseases and conditions. However, developments in the field of molecular biology have led to an increase in the knowledge of biological systems and their interactions. Proteins are biomolecules that are essential in determining the structure and carrying out most of the functions in living cells. The future of pharmacy belongs to a special category of protein therapeutics. Based on the result of industry analysis, it is suggested how to commercialize protein therapeutics for biotech-enterprises.

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Construction of Genetic Linkage Map using Microsatellite and SNP Markers in Korean Native Chicken (Microsatellite와 SNP Marker를 이용한 한국재래닭의 유전적 연관지도 작성)

  • Seo, Dong Won;Park, Hee Bok;Choi, Nu Ri;Jin, Shil;Yoo, Chae Kyoung;Sultana, Hasina;Heo, Kang Nyeong;Jo, Cheorun;Lee, Jun Heon
    • Korean Journal of Poultry Science
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    • v.42 no.1
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    • pp.77-86
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    • 2015
  • Chicken is one of the major livestock, especially for supplying proteins to human. The chicken genome size is approximately one-third compared with that of the human genome and regarded as a valuable model animal for genetics and development biology. In this study, we constructed the genetic linkage map for Korean native chicken (KNC) using 131 microsatellite (MS) and 8 single nucleotide polymorphism (SNP) markers. As a result, the total map length was calculated as 2729.4 cM and the average genetic distance between markers was 19.64 cM. The marker orders and genetic distances were well matched with the consensus linkage map except for the physical order of ADL0278 and MCW0351 in GGA8. In addition, the recombination rates in marcrochromosomes were 3.7 times higher than that of microchromosomes. The average numbers of alleles, expected heterozygosity (Hexp) and polymorphic information content (PIC) values were calculated as 5.5, 0.63 and 0.58, respectively. These results will give useful information for the understanding of genetic structure and QTL studies in KNC.