• Title/Summary/Keyword: DNA-DNA hybridization

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Hot Pepper Functional Genomics: Monitoring of Global Gene Expression Profiles During Non-Host Resistance Reactions in Hot Pepper Plant ( Capsicum annuum).

  • Lee, Sanghyeob;Chung, Eun-Joo;Park, Doil
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.80.2-81
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    • 2003
  • Since hot peppers (Capsicum annuum L.) are getting reputation as an important source of vitamins, medicine and many other areas, consumption and cultivation is being increased in the world. In spite of this usefulness, so little attention has been given to the hot pepper plants. To date, less than 500 nucleotide sequences including redundancy has been identified in NCBI database. Therefore we started to EST sequencing project for initial characterization of the genome, because of the large genome size of hot pepper (2.7 3.3 ${\times}$ 109 bp), To date, a set of 10,000 non-redundant genes were identified by EST sequencing for microarray-based gene expression studies. At present, cDNA microarrays containing 4,685 unigene clones are used for hybridization labeled targets derived from pathogen infected and uninoculated leaf tissues. Monitoring of gene expression profiles of hot pepper interactions with soybean pustule pathogen (Xag;Xanthomonas axonopodis pv. glycine) will be presented.

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Aspergillus niger로 부터 $\alpha$-glucosidase 발현억제 형질전환체의 분리

  • 이동건;이진영;서영배
    • Microbiology and Biotechnology Letters
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    • v.24 no.4
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    • pp.427-429
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    • 1996
  • We have already cloned an extracellular $\alpha$-glucosidase gene from Aspergillus niger with oligonucleotide probe synthesized on the basis of the peptide sequences determined previously. The DNA sequence revealed an open reading frame of 895 amino acids split by three introns. We are attempting to construct an A. niger strain deficient in the $\alpha$-glucosidase enzyme activity, which would be useful for the glucoamylase production without contamination by the industrially undesirable $\alpha$-glucosidase. For destruction of the $\alpha$-glucosidase gene, we try to make transformations. A cloned partial $\alpha$-glucosidase gene was introduced into Aspergillus niger, and transformants with suppressed $\alpha$-glucosidase activity were isolated. The transformants were cultured on YPD medium which contained Hygromycin B at 30$\circ$C. The activity of $\alpha$-glucosidase of the suppressed transformants was compared to that of wild type activity. As shown by southern-hybridization, we detected that the transformant was a heterocaryon.

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A Genome-Specific PCR Primer Design Program for Open Reading Frames

  • Keong, Kwoh-Chee;Lim, Kok-Wui
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.147-150
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    • 2005
  • Proper PCR primer design determines the success or failure of Polymerase Chain Reaction (PCR) reactions. In this project, we develop GENE-PRIMER, a genomes specific PCR primer design program that is amenable to a genome-wide scale. To achieve this, we incorporated various parameters with biological significance into our program, namely, primer length, melting temperature of primers Tm, guanine/cytosine (GC) content of primer, homopolymeric runs in primer and self-hybridization tendency of primer. In addition, BLAST algorithm is utilized for the purpose of primer specificity check. In summary, selected primers adhered to both physico-chemical criteria and also display specificity to intended binding site in the genome.

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Molecular cloning and nucleotide sequence of schizosaccharomyces pombe Homologue of the receptor for activated protein kinase C gene

  • Park, Seung-Keil;Yoo, Hyang-Sook
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.128-131
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    • 1995
  • Using differential hybridization, we selected the prk gene fortuitously from Schizosaccharomyces pombe homologous to RACK1 of rat which encodes the receptor for activated protein kinase C. The cDNA sequence of prk was determined and its deduced amino acid sequence was 76% homologous to RACK1 and had the feature of trimeric G protein bata subunit. The specific amino acid sequences required for the protein kinase C binding were also present in Prk as in the case of RACK1 protein. From these similarities, we suggest that the Prk is protein kinase C binding protein of S. prombe. The involvement of Prk in signal transduction mediated by protein kinase C remained to be studied.

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수계 생태계에서의 세균 군집 구조의 분자생물학적 분석

  • 이동훈;김상종
    • Korean Journal of Microbiology
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    • v.33 no.1
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    • pp.55-65
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    • 1997
  • 16S rRNA를 분석한 연구들은 자연 생태계에서 추출한 핵산을 이용하여 16rRNA 유전자의 염기서열을 분석하거나 특정 DNA probe를 이용한 hybridization 실험이 주류를 이루어 왔다. 특히 PCR 기법이 개발됨에 따라 적은 양의 시료를 대량으로 손쉽게 증폭시킬 수 있어 다양한 분야에 응용되고 있다. 세균 군집의 구조를 이해하는데 있어서 PCR 방법의 적용 대상은 주로 16S rRNA 유전자의 염기서열 해독분야이며 해양 생태계를 대상으로 많은 연구 결과가 보고되었다(11,13,21,26). 한편 자연 생태계의 개별적 미생물 분류룬들을 검출하기 위한 특정 oligonucleotide probe의 개발방법들은 미생물 군집의 유전적 다양성에 대한 정보 파악 이외에 배양이 어려운 혐기성 세균과 같은 특정 세균들의 동정에도 이용되고 있다(3,24,55). 본고에서는 세균 군집의 구조와 다양성을 연구하는데 적용 가능한 rRNA 분석방법들을 수계 생태계를 중심으로 살펴보고자 한다.

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Production of a Normal Calf from Bovine Embryo Microinjected with Human Growth Hormone Gene (사람 성장호르몬 유전자가 미세주입된 체외수정란 유래의 송아지 생산)

  • 손동수;김선정;김일화;서국현;이광원;상병돈;박무균;이철상;한용만
    • Journal of Embryo Transfer
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    • v.9 no.3
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    • pp.229-234
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    • 1994
  • This experiment was carried out to develop the model system for mass production of biomedical and nutritional proteins (human proteins) through mamraary gland of the transgenic cattle produced by gene manipulation and embryological technologies. Human growth hormone gene fused with rat $\beta$-casein gene promoter was microinjected into pronuclei of one cell bovine embryos produced by in vitro fertilization. After microinjection, embryos were cultured in vitro for 6 or 7 days. Twenty embryos reaching to blastocysts were transferred to 10 beef recipients, each receiving two embryos. Recipients were diagnosed for pregnancy by rectal palpation at 76 days after embryo transfer. One of them was pregnant to term and produced a female calf weighing 21 kg at 280 days following embryo transfer. DNA was extracted from umbilical cord tissue and blood of calf born for confirming gene insertion. As determined by Southern hybridization, the transgene was not found.

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Molecular Cloning of Antagonistic Genes in Pseudomonas maItophiliQ B-14 (토양병해 길항성 Pseudomonas maltophilia B-14의 길항유전자탐색)

  • 구본성;서영우;윤상홍;박경수;은무영;김용환;오상우;류진창;은무영
    • Microbiology and Biotechnology Letters
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    • v.20 no.6
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    • pp.619-624
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    • 1992
  • Tn5 lac 삽입으로 채소입고병원균에 길항력이 약화된 T-67 및 고추역병균과 참깨역병균에 길항력이 약화된 T-81의 Tn5 lac 유전자 일부와 오른쪽 말단에 있는 길항관련 유전자의 flanking sequence가 cloning된 pAG67 및 pAG81 clone을 선발하였고, pAG67 및 pAG81 clone된 길항관련 유전자의 flanking sequence를 야생 길항균 Pseudomonas maltophilia B-14의 DNA를 probe로 사용하여 Southern hybridization으로 확인하였으며, 제한효소 지도를 작성하여 8Kb 및 4Kb 크기의 flanking sequence가 cloning되었음을 확인하였다. pAG6 및 pAG81의 flanking sequence를 EcoRi-BglII와 EcoRI-MpaI으로 분리하여 유전자 은행으로부터 길항관련 유전자가 cloning된 cosmid clone 7개주를 선발하였다.

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Telomere Erosion-Induced Mitotic Catastrophe in Chinese Hamster Don Cells

  • Sohn, Sea H.;Multani, Asha S.;Gugnani, Pankaj K.;Pathak, Sen
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.15-15
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    • 2002
  • Telomere erosion is the earliest chromatin modification in cells entering the apoptotic regime. The purpose of this investigation was to demonstrate that loss of telomeric DNA was involved in the induction of mitotic catastrophe and cell death of Chinese hamster Don cells that were grown continuously without subculturing for several days. Don, which is a diploid male Chinese hamster-derived cell line, was grown without sub-culturing for one to four days at 37℃ and then harvested for cytological preparations. (omitted)

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Parthenogenesis in Silkworm, Bombyx mori L.

  • Gangopadhyay Debnirmalya;Singh Ravindra;Kariappa B. K.;Dandin S. B.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.1
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    • pp.1-10
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    • 2005
  • Parthenogenesis in mulberry silkworm, Bombyx mori L. acquires immense use in the development of outstanding homozygous lines with higher viability, hybrid vigour, combining ability and less phenotypic variability. It can serve as a powerful tool in controlling sex of the offsprings as well as a useful tool in selection. In fact India is the second largest silk producing country in the world next only to China and all the five types of natural silks viz., mulberry, oak tasar, tropical tasar, muga and eri are produced in India. However, little information is available on the role of artificial parthenogenesis in the development of superior silkworm breeds. This paper overviews some important studies carried out on artificial parthenogenesis, and outline of different types of parthenogenesis, methods of induction of artificial parthenogenesis, factors responsible for successful parthenogenetic development, cytogenetics of artificial parthenogenesis and role of artificial parthenogenesis in silkworm breeding. Besides, an attempt is made to describe briefly about parthenogenetic engineering which includes cloning in silkworm, artificial insemination, chimeras, hybridization, chromosomal substitution and recombinant DNA in silkworm.

형질전환 연초의 복합바이러스 저항성

  • 이기원;채순용;이청호;이영기;강신웅;박성원;박은경
    • Journal of the Korean Society of Tobacco Science
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    • v.21 no.1
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    • pp.70-76
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    • 1999
  • KF 116 was TMV resistant tobacco plant and KB 301 was PVY resistant plant transformed with TMV CP gene and PVY CP gene, respectively. These resistant plants were cross-fertilized and the 4 lines of the TMV-PVY resistant plants were selected from F1 hybrid plants. The rate of PVY-resistant plant in these hybrids was 100 percent and that of TMV-resistant plants including delay type was 90-98 percent at 4 weeks after virus inoculation. It was confirmed that the TMV and PVY CP genes were integrated into the genome of hybrid plants by genomic PCR, and Southern blot hybridization. The genome of F1 hybrid plants had one copy and 4 copies of PVY-CP gene and TMV-CP gene, respectively, and CaMV 35S promoters were not methylated, regardless of the difference symptom development to TMV.

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