• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.838-843
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• 2005

Brain-derived neurotrophic factor (BDNF) is a small secretory protein and a member of the nerve growth factor (NGF) gene family. We cloned the flounder BDNF gene from a flounder brain cDNA library. The nucleotide sequence of the cloned gene showed an open reading frame (ORF) consisting of 810 bp, corresponding to 269 amino acid residues. The tissue distribution of flounder BDNF was determined by reverse transcription-polymerase chain reaction (RT-PCR) in brain, embryo, and muscle tissues. To express fBDNF using a eukaryotic expression system, we constructed the vector mpCTV-BDNF containing the fBDNF gene and transformed this vector into Chlorella ellipsoidea. Stable integration of introduced DNA was confirmed by PCR analysis of genomic DNA, and mRNA expression in C. ellipsoidae was confirmed by RT-PCR analysis.

• Archives of Pharmacal Research
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• 제13권1호
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• pp.69-73
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• 1990

As a part of the study to elucidate the mechanism by which transcription of LDH A-gene is regulated by cAMP, we aimed to isolated rat LDH A gene and characterize cAMP-reponsive element (CRE). We have screened $1.2{\times}10^6$ recombinant phages of rat Charon 4A genomic library and isolated 33 positive clones among which we identified 12 different LDH A gene-related clones. By the results of restriction enzyme mapping, Southern blotting, and nucleotide sequence analyses, we concluded that the 12 LDH A gene-related clones were intronless and frequently mutated LDH A-pseudogenes. In this report, we present the characteristic features of the 12 rat liver LDH A-pseudogenes.

• Mycobiology
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• 제30권2호
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• pp.88-92
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• 2002

Rehmannia glutinosa is a perennial medicinal plant belonging to the family Scrophulariaceae with more than 300 species known in the world, especially in temperate regions. Its roots have been used widely in Korea for medicinal purposes. However, it is commonly infected by various pathogens during storage, causing great damage to the roots, and impedes the intensive farming of the crop. Therefore, an attempt has been made to isolate and screen a resistance gene against the pathogen Fusarium oxysporum using differential display. We treated salicylic acid(SA), and isolated a resistance gene that responds to SA. As a result, we found that SA was involved in plant defense mechanism in pathogenicity tests with SA treated and non-treted plants, and we isolated a partial PR-la gene through differential display polymerase chain reaction(DD-PCR) method.

• 한국동물학회지
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• 제37권1호
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• pp.12-18
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• 1994

The prothoracicotropic hormone (PTTH) produced by the neurosecretory cells in insects is involved in molting and metamorphosis by activating the prothoracic frins) glands to secrete ecdysone (or related ecdvsteroidsl. In the present study, the small PTTH-like gene was isolated by screening of CDNA library using the bombvxin (corresponding to small PTTH in Bombvx moril gene probe in Drosophilo melonogaster. It showed 50-6096 sequence homology to bombyxin gene. The expression patterns of this gene showed developmental stage- and tissue-dependent manners. The mRNA was detected only in the late third instar larval-prepupa which is stases showing the highest hormonal activity to secrete ecdysteroids, and detected in the brain pan of the Isle third instar lanrae.

• 미생물학회지
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• 제26권1호
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• pp.32-36
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• 1988

The yeast gene HOM3 encodes aspartokinase, which catalyses the first step (aspartate to and from beta-aspartyl phosphate) of common pathway to threonine and methionine. The yeast HOM3 gene expression is known to be regulated by threonine and methionine specific control, and also by general control of amino acid biosynthesis. Isolation and characterization of the HOM3 gene are essential for the molecular genetic study on its regulation of expression. A recombinant plasmid pSC3 (15.5kb, vector YCp50) has been cloned into E. coli HB101 from yeast genomic library through their complementing activity of HOM3 mutation in a yeast recipient strain M34-24B. Organization of the plasmid was characterized by delineation of restriction cleavage sites in the insert fragment.

• 한국식품영양학회지
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• 제5권1호
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• pp.41-48
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• 1992

Megaplasmids of Azospirillum strains isolated in the Korean paddy field were identified. Five megaplasmids were identified from Azospirillum lipoferum AS192. Homology between nod ABC gene of Rhizobium meliloti and megaplasmids of Azospirillum lipoferum AS192 and Azospirillum brasilense AS112 was found. This observation might have reflected a common mechanism in the early process of soil bacterial association with plants.

• 한국작물학회지
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• 제51권spc1호
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• pp.274-276
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• 2006

다량의 페놀류 2차 대사산물과 다당류가 포함된 녹두 종자의 RNA를 Tris-buffer와 guanidium thiocyanate 용액을 이용하여 손쉽고 간편하게 고순도의 RNA를 분리하는 방법을 확립하였으며, 우리의 분리 방법을 이용하여 추출한 RNA를 사용하여 RT-PCR과 유전자 클로닝 등의 분석시험에 이용할 수 있었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.527-530
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• 2000

중국산 야생키위로 total RNA를 추출하고 RT-PCR를 실시해서 증폭된 1.2kb fragment를 얻어 pGEM-T Easy에 cloning한 후 염기서열과 아미노산 서열을 비교 분석한 결과 actinidin gene인 것으로 분석되었다.

• Journal of Microbiology and Biotechnology
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• 제4권1호
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• pp.20-23
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• 1994

The polymerase chain reaction was used to study the genes inducible under stress from the heavy metal cadmium. Schizosaccharomyces pombe, grown in the presence or absence of sublethal concentration of cadmium, was isolated to purify the total RNAs. The Induced RNA Random Fishing (IRRF) method in which random oligonucleotides were used as primers was applied to the identification of cadmium-induced gene expressions. A PCR-DNA product of 400-bp was cloned and sequenced. Computer analysis showed that this DNA has no homology with any known DNA sequences in GenBank or EMBL databases. The induction of this gene was confirmed by Northern blot analysis of total RNAs isolated from both cadmium-treated and untreated yeast cells.

• 미생물학회지
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• 제30권6호
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• pp.488-494
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• 1992

Three clones containing mouse CD4 gene were prepared using AKR genomic cosmid library. The role of 6, 500 bp 5' flanking region of the first exon of the AKR CD4 gene in tissue or developmental stage specific expression of CD4 has been studied. The deletion constructs containing various amounts of CD4 5' flanking sequences were prepared, and they were transfected into the cell lines representing different cell types or developmental stages of CD4 expression. Study of the reporter gene expression revealed that at least 1, 700 bp of 5' flanking region did retain promoter activity for CD4 expression. This area did not seem to contain enhancer activity for a full expression of CD4. However, the putative promoter interacted with other tissue specific enhancer sequence and showed the tissue specificity of the enhancer element.