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BcHSP17.6 유전자 도입에 의한 알팔파의 형질전환 (Transformation of Alfalfa by BcHSP17.6 Gene using Agrobacterium tumefaciens)

  • 김기용;성병렬;임용우;최기준;임영철;장요순;서성;윤세형;박근제;조진기
    • 한국초지조사료학회지
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    • 제21권3호
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    • pp.151-156
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    • 2001
  • 내열성 유전자인 BcHSP17.6를 갖도록 제작한 발현벡터 pBKH4를 Agrobacterium tumefaciens LBA 4404에 도입후, Agrobacterium과 알팔파 캘러스의 공배양을 통해 감염시킨 캘러스를 $100{\mu}g/m{\ell}$의 kanamycin과 $500{\mu}g/m{\ell}$의 cefotaxim을 첨가한 SH-kc배지에서 배양하며 형질전환된 캘러스를 선발하였다. 식물체 재분화는 SH- nk-c, SH-sp-c, SH-11b-c, SH-1BA 배지에서 약 4개월간 배양하여 재분화를 완성하였으며, 재분화된 알팔파의 genomic DNA를 분리한 후, PCR 분석 및 Southern blot 분석을 실시하여 알팔파의 형질전환을 확인하였다.

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Development of Artificial Insemination Techniques with a Minimum Numbers of Insemination Spermatozoa using Laparoscopy

  • Lee, J. H.;Park, S. J.;I. S. Ryu;G. Y. Chung;Park, S. H.;D. Y. Ji;Kim, C. K.;S. H. Baek
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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    • pp.250-250
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    • 2004
  • This study was conducted to investigate the effect of the number of spermatozoa and insemination section(field) of reproductive organs at artificial insemination using laparoscopy(Fiber optic laparoscopic system, Good-Gene Co., Korea) in deer(Elk) and cattle. Twenty six elk does and fifteen cows were inserted CIDR into virginia during 12∼14 days for synchronization of estrus. (omitted)

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Expression of GFP Gene in Porcine Embryos after ICSI with Different DNA Binding Methods

  • Han, J. H.;Kim, S.W.;Lee, Y.K.;Lee, P.Y.;Park, C.G.;Lee, S.E.;Baek, K.N.;Lee, H.G.;Lee, J.Y.;Chang, W.K.;Park, J.K.
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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    • pp.191-191
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    • 2004
  • Transgenic animals are produced primarily by microinjecting exogenous DNA into the male pronuclei of a zygote. Microinjection method for gene transmitting is successful in mice but not efficient in farm animals, limiting it's general utility such as a large scale facility and labour. Based on our finding that sperm cells bind with exogenous DNA, sperm was used as a vector for producing transgenic animals to introduced green fluorescence protein(GFP) gene. (omitted)

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Characterization of Lupinus Iuteus Chloroplgsl Gene Coding for Components of a Chloroplastic NADH Dehydrogenase

  • Oczkowski, Marian;Augustyniak, Halina
    • Journal of Plant Biotechnology
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    • 제2권2호
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    • pp.73-78
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    • 2000
  • The plastid genomes of several plants contain ndh genes homologues of genes encoding subunits of the mitochondrial complex I. We sequenced the part of lupin ndhB, ndhD and ndhF genes in order to compare the structure of these genes with those of Nicotiana tabaum, Arabidopsis thaliana, Zea mays and Oryza sativa with the idea to detect the presence of stretches with identical aminoacid composition. We were only able to find one or two stretches of this kind of about 16 aminoacid- long in the analyzed fragments of the ndh genes. The total number of such stretches was different in particular gene products: for ndhc 1, ndhB 9, ndhD 3 and ndhF 6. We have also examined the transcription pattern of ndhC, ndhK and ndhJ genes during lupin development. We show that the greatest amount of ndhC, ndhK and ndhJ transcripts are observed in 7- to 14 day- old lupin seedlings. We also studied the level of transcription of those genes in plants growing at low temperature. All the data confirmed that the abundance of transcription of ndhC, ndhK, and ndhJ genes increased under chill conditions. It has to be noted that the level of transcription of the ndhC gene was higher than the other genes probably due to higher stability of this transcript.

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전산생물학을 이용한 마이크로어레이의 유전자 발현 데이터 분석 및 유형 분류 기법 (Analysis and Subclass Classification of Microarray Gene Expression Data Using Computational Biology)

  • 유창규;이민영;김영황;이인범
    • 제어로봇시스템학회논문지
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    • 제11권10호
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    • pp.830-836
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    • 2005
  • Application of microarray technologies which monitor simultaneously the expression pattern of thousands of individual genes in different biological systems results in a tremendous increase of the amount of available gene expression data and have provided new insights into gene expression during drug development, within disease processes, and across species. There is a great need of data mining methods allowing straightforward interpretation, visualization and analysis of the relevant information contained in gene expression profiles. Specially, classifying biological samples into known classes or phenotypes is an important practical application for microarray gene expression profiles. Gene expression profiles obtained from tissue samples of patients thus allowcancer classification. In this research, molecular classification of microarray gene expression data is applied for multi-class cancer using computational biology such gene selection, principal component analysis and fuzzy clustering. The proposed method was applied to microarray data from leukemia patients; specifically, it was used to interpret the gene expression pattern and analyze the leukemia subtype whose expression profiles correlated with four cases of acute leukemia gene expression. A basic understanding of the microarray data analysis is also introduced.

한국 재래닭의 경제형질 개량을 위한 apoVLDL-II 유전자의 유전자형 분석 (Genotype Analysis of apoVLDL-II Gene in Korean Chicken Breeds)

  • 정기철;이유주;;장병귀;최강덕;이준헌
    • 한국가금학회지
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    • 제35권4호
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    • pp.335-339
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    • 2009
  • 초저밀도 아포지단백(Very Low Density Apolipoprotein-II)은 조직의 지방단백질 분비 조성과 밀접한 관련이 있다고 알려진 유전자로서 최근 닭에서 성장 및 체구성과 매우 높은 연관이 있다고 알려져 있다. 본 연구는 닭의 3품종에서 PCR-RFLP 방법을 통해 apoVLDL-II 유전자의 유전자형을 분석하고 Broiler 형(B)과 Fayoumi 형(F)을 결정하였다. 각 품종간 유전자형 빈도는 한국 재래계에서 BB가 0.37, BF가 0.43, FF가 0.2로 검출되었고, 오계에서 BB가 0.2, BF가 0.6, FF가 0.2로 검출되었으며, 백색레그혼에서 BB가 0.16, BF가 0.84로 검출되었다. Broiler 형은 성장이나 체구성을 높이는 방향으로 작용한다는 기존의 보고를 바탕으로 본 연구에서 나타난 집단내 다양한 유전자형 분포는 재검증 작업을 거쳐 육종에 의해 이 형질들을 개량할 수 있음을 의미한다.

Extension of a 5'- or 3'-end Genomic DNA Sequence by a Single PCR Amplification

  • Jeon, Taeck J.
    • 통합자연과학논문집
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    • 제1권3호
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    • pp.230-233
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    • 2008
  • A simple and rapid method is described for extending the 5'- or 3'-end genomic sequence of a known partial sequence by only a single round of PCR. This method involves digesting and ligating genomic and plasmid DNAs, and amplifying the 5'-upstream or 3'-end downstream sequence of the known DNA sequence, using two primers, one gene specific and the other plasmid specific. A single round of PCR amplification is sufficient to produce gene-specific bands detectable in gels. By using this approach, 5'-end genomic sequence of the D-amoeba sams gene was extended.

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Cloning and Strong Expression of a Bacillus subtilis WL-3 Mannanase Gene in B. subtilis

  • Yoon, Ki-Hong;Lim, Byung-Lak
    • Journal of Microbiology and Biotechnology
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    • 제17권10호
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    • pp.1688-1694
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    • 2007
  • A gene encoding the mannanase of Bacillus subtilis WL-3, which had been isolated from Korean soybean paste, was cloned into Escherichia coli and the nucleotide sequence of a 2.7-kb DNA fragment containing the mannanase gene was subsequently determined. The mannanase gene, designated manA, consisted of 1,080 nucleotides encoding a polypeptide of 360 amino acid residues. The deduced amino acid sequence was highly homologous to those of mannanases belonging to glycosyl hydrolase family 26. The manA gene was strongly expressed in B. subtilis 168 by cloning the gene downstream of a strong B. subtilis promoter of plasmid $pJ27{\Delta}88U$. In flask cultures, the production of mannanase by recombinant B. subtilis 168 reached maximum levels of 300 units/ml and 450 units/ml in LB medium and LB medium containing 0.3% locust bean gum, respectively. Based on the zymogram ofthe mannanase, it was found that the mannanase produced by recombinant B. subtilis could be maintained stably without proteolytic degradation during the culture time.

Xanthomonas sp. YL-37의 Alkaline Protease 유전자의 클로닝 (Cloning of a Alkaline Protease Gene from Xanthomonas sp. YL-37)

  • 이대희;김수경;이승철;윤병대;황용일
    • 한국미생물·생명공학회지
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    • 제23권2호
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    • pp.145-149
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    • 1995
  • For the purpose of developing a new biodegradable detergent, we have isolated a gene encoding wide-range temperature applicable alkaline protease from Xanthomonas sp. YL-37 (Lee et al., 1994, Kor. J. Appl. Microbiol. Biotechnol.). An alkaline protease gene was isolated from the gene bank that was prepared from the chromosomal DNA of Xanthomonas sp. YL-37. From the results of agarose gel electrophoresis and a restriction enzyme mapping, a 2.7 kb DNA fragment containing the alkaline protease gene was inserted in the plasmid pUC9. Extracellular activity of a clone having alkaline protease gene was detected on SDS-polyacrylamide gel with activity staining assay. The molecular weight of alkaline protease was determined to be about 64 kDa from 11% SDS-PAGE analysis. Alkaline protease activity, produced from E. coli which harboring the plasmid, showed no difference at reaction temperature 20, 30 and 40$\circ$C, respectively. This result showed that alkaline protease produced from E. coli harboring the plasmid was apparently the same as that of Xanthomonas sp. YL-37.

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