• 대한의생명과학회지
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• 제6권3호
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• pp.165-170
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• 2000

Vacuolar (H$^{+}$)-ATPase (V-ATPase)는 multi-subunit로 구성된 단백질로서, proton pumping을 통해 세포내 산성화반응에 관여를 한다. 최근에 이 단백질이 synaptic vesicle에서도 발견된 것으로 보아 뇌 신경전달에 중요한 역할을 수행할 것으로 추정하고 있다. 우리는 흰쥐 뇌에서 분리한 mRNA를 주형으로 한 PCR 반응에서 16 kDa subunit의 V-ATPase cDHA를 클로닝할 수 있었고, 이의 염기서열 또한 결정하였다. 분리된 뇌 16 kDa V-ATPase의 coding sequence는 전체 468 bp로서 간에서 보고되었던 것과 동일한 크기였다. 단지 3' 말단의 염기 하나가 A에서 C로 바뀌어 있었는데 모두 alanine (GCA, GCC)을 지정하기 때문에 단백질의 일차구조에는 변화가 없는 것으로 확인되었다. 한편 rat brain cDNA library에서도 동일한 clone이 분리되었는데 역시 같은 부분에서 polymorphism이 발견되었고, RNA splicing 등 더 이상의 조직특이적 변화는 없었다. 본 연구는 16 kDa V-ATPase의 뇌에서의 기능과 신경말단에서의 neurotransmission 및 synaptic vesicle의 재순환 기전을 이해하는데 유용한 정보가 될 것이다.

• ALGAE
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• 제37권1호
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• pp.1-14
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• 2022

Cyanobacteria are distributed worldwide, and many new cyanobacterial species are discovered in tropical region. The Nostoc-like genus Amazonocrinis has been separated from the genus Nostoc based on polyphasic methods. However, species diversity within this genus remains poorly understood systematically because only one species (Amazonocrinis nigriterrae) has been described. In this study, two novel strains (NUACC02 and NUACC03) were isolated from moist rice field soil in Thailand. These two strains were characterized using a polyphasic approach, based on morphology, 16S rRNA phylogenetic analysis, internal transcribed spacer secondary structure and ecology. Phylogenetic analyses based on 16S rRNA gene sequences confirmed that the two novel strains formed a monophyletic clade related to the genus Amazonocrinis and were distant from the type species A. nigriterrae. The 16S rRNA gene sequence similarity (<98.1%) between novel strains and all other closely related taxa including the Amazonocrinis members exceeded the cutoff for species delimitation in bacteriology, reinforcing the presence of a new Amazonocrinis species. Furthermore, the novel strains possessed unique phenotypic characteristics such as the presence of the sheath, necridia-like cells, larger cell dimension and akinete cell arrangement in long-chains and the singularity of D1-D1', Box-B, V2, and V3 secondary structures that distinguished them from other Amazonocrinis members. Considering all the results, we described our two strains as Amazonocrinis thailandica sp. nov. in accordance with the International Code of Nomenclature for Algae, Fungi and Plants.

• ALGAE
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• 제38권2호
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• pp.93-110
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• 2023

Progress in phylogenomic analysis has led to a considerable re-evaluation of former cyanobacterial system, with many new taxa being established at different nomenclatural levels. The family Geminocystaceae is among cyanobacterial taxa recently described on the basis of polyphasic approach. Within this family, there are six genera: Geminocystis, Cyanobacterium, Geminobacterium, Annamia, Picocyanobacterium, and Microcrocis. The genus Geminocystis previously encompassed two species: G. herdmanii and G. papuanica. Herein, a new species G. urbisnovae was proposed under the provision of the International Code of Nomenclature for algae, fungi, and plants (ICN). Polyphasic analysis was performed for five strains from the CALU culture collection (St. Petersburg State University, Russian Federation), and they were assigned to the genus Geminocystis in accordance with high 16S rRNA gene similarity to existing species, as well as because of proximity to these species on the phylogenetic trees reconstructed with RaxML and Bayes methods. Plausibility of their assignment to a separate species of the genus Geminocystis was substantiated with smaller cell size; stenohaline freshwater ecotype; capability to complementary chromatic adaptation of second type (CA2); distinct 16S rRNA gene clustering; sequences and folding of D1-D1' and B box domains of the 16S-23S internal transcribed spacer region. The second objective pursued by this communication was to provide a survey of the family Geminocystaceae. The overall assessment was that, despite attention of many researchers, this cyanobacterial family has been understudied and, especially in the case of the crucially important genus Cyanobacterium, taxonomically problematic.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.323-323
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• 2022

Jack bean [Canavalia ensiformis (L.)], belonging to the Leguminosae family has been frequently used in edible and medicinal plants in Asian countries. Jack beans are high in protein which is approximately 30%. Concanavalin A (Con A) is a major protein of Jack bean and belongs to the family of legume lectins. It has inhibitory effect on hepatocellular carcinoma by inducing autophagy. However, Con A negatively affects nutrient utilization by other mechanisms. It binds to the glycoproteins and glycolipids of the digestive tract mucosa, inhibits the activity of the enzymes of the brush border of the enterocytes. In order to use Jack bean young seedpods, they are restricted to 'young pods (soft, pre-swelling)' according to the 'Food Code' (Ministry of Food and Drug Safety). Therefore, in this study, we investigated the quantitative change of Con A across developmental stages of Jack bean pods. Biological samples consisted of Jack bean pods and seeds in 7 stages of development. The expression pattern of Con A mRNA was monitored by quantitative reverse transcription PCR (RT-qPCR). Expression of Con A proteins was analyzed by western blotting. The expression of Con A mRNA and protein in the seeds tended to increase gradually as the seeds expanded. However, in pods, they were much less than in seeds. As the expression of Con A mRNA and protein increases as the pods thicken, it is predicted that Con A synthesis increases when the thickness growth of the pod begins after the length growth of the pod is completed. Since the expression of Con A in the pods and seeds in very low when the pods are about 2 cm, therefore 2 cm pods seem appropriate when using 'young pods'. It is also necessary to study other proteins in Jack bean, such as Urease and Canavalin. These studies will serve as the basis for processing Jack bean.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.607-607
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• 2003

• 대한유전성대사질환학회지
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• 제5권1호
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• pp.108-115
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• 2005

Inherited metabolic diseases (IMD) comprise a large class of genetic diseases involving disorders of metabolism. The majorities are due to defects of single genes that code for enzymes that facilitate conversion of various substances into others. Because of the multiplicity of conditions, many different diagnostic tests are used for screening of IMD. Molecular genetic diagnosis is the detection of pathogenic mutations in DNA and/or RNA samples and is becoming a much more common practice in medicine today. The purpose of molecular genetic testing in IMD includes diagnostic testing, pre-symptomatic testing, carrier screening, prenatal diagnosis, preimplantation testing, and population screening. However, because of the complexity, difficulty in interpreting the result, and the ethical considerations, an understanding of technical, conceptual, and practical aspects of molecular genetic diagnosis is mandatory.

• 한국미생물·생명공학회지
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• 제13권1호
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• pp.19-25
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• 1985

Saccharomyces cerevisiae HIS 5 유전자는 histidinol phosphate aminotransferase (EC: 2, 6, 1,9)를 code하는 아미노산 합성유전자이다. 이 유전자는 plasmid pSH 530에 cloning되어 E. coli와 Saccharomyces cerevisiae 숙주에서 promoter로서 전사하였다. HIS 5 유전자의 총염기 수는 736개이였고 5' 상류영역에는 긴 reading frame, directed repeat, 전사개시점, 그리고 Pribnow box염기배열이 있었다. 특히 HIS 5 유전자의 ATG 주변 염기배열은 -A-A-A-T-T-A-C-A-C-T-A-T-G-G-T-T-T-T-T-G-A-T-였으며 C block은 없었다.

• 생명과학회지
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• 제24권6호
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• pp.694-699
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• 2014

토착 미생물은 친환경적 정화에 중요한 역할을 한다. 원유(crude oil)를 분해하는 80균주를 인천 연안에서 분리하고 oil film collapsing방법을 이용하여 유화능이 있는 12균주를 선별하였다. 이들 균주에 대해 ${\rho}$-nitrophenyl butylrate를 기질로 이용하여 리파아제(lipase)활성과 n-hexanedecane을 기질로 이용하여 유화(emulsification)활성을 측정하여 원유 분해 활성이 좋은 Incheon9를 선별하고 gas chromatography (FID)로 paraffine계 탄화수소를 감소시키는 것을 확인하였다. 이 균주의 16s rRNA유전자 분석을 통해 Acinetobacter sp.로 동정하고 NCBI에 등록하여 accession code (KF548540)를 부여 받았다. Acinetobacter sp. Incheon9의 성장과 유화능이 최적 배양 조건은 $20^{\circ}C$, pH 7, 1% NaCl였으며 대수증식기 기간에 가장 높은 유화능을 보였으며 탄화수소가 짧은 trybutyrin에서 분해능력이 좋았다. 이번 연구결과는 환경오염에 활용 가능한 미생물자원군의 확보를 위한 연구였으며 추후 활용을 위해서는 실제 환경에서 동일한 활성을 가지는지 여부에 대한 연구가 추가로 진행되어야 할 것이다.

• 한국토양비료학회지
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• 제33권4호
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• pp.266-274
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• 2000

토양과 작물근계의 유용미생물을 이용하여 작물생산성을 증대하고 병충해의 생물학적 방제를 위해서는 토양-근계의 미생물군집을 분석하고 그 기능을 밝히는 것이 전제가 되어야한다. 그러나 희석평판법으로는 극히 일부분만이 배양된다는 점을 고려할 때, 생존하지만 배양 불가능한 미생물의 군집 분석도 반드시 병행할 필요가 있다. 따라서 본 연구에서는, 고추재배지의 토양, 근권토양, 근면의 세균군집 구조해석을 위해서, 배양을 거치지 않고 각 시료로부터 직접 DNA를 추출하여 PCR증폭, 16S rDNA cloning, sequencing, 계통 해석을 행했다. 그 결과, 토양중에는 근권세균보다 미지의 동정이 되지 않는 세균이 우점하고 있었다. 27 clones 중에서 16 clones이 그램음성세균의 대표격인 Proteobacteria였으며, 방선균 등이 속해있는 고(高) G+C 그램양성세균군은 1 clone이 검출되었다. 그 외는 CFB 군이 2 clones, Verrucomicrobia가 1 clone이었고, Nitrospira가 1 clone이었으며 4 clones은 어느 군에도 속하지 않았다.

• 한국미생물·생명공학회지
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• 제22권2호
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• pp.134-142
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• 1994

The neucleotide sequences of the xylA gene encoding $\beta $-xylosidase of Bacillus stearothermophilus and is its flanking regions were datermined. Three open reading frame(ORFs) were found, one of which(ORF1) appeared to code for the $\beta $-xylosidase. The 1830 base pair ORF1 encoded 609 amino acids starting from a TTG initiation codon. The molecular weight deduced from the nucleotide sequence(68 KD) was in agreement with that estimated by SDS-polyacrylamide gel electrophoresis of the purified enzyme(66 KD). The Shine-Dalgarno sequence(5'-AGGAGG-3') was found 11 bp upstream of the initiation codon. Further 15 bp upstream, there observed a potential transcription initiation signals. The putative -10 sequence(CATAAT) and -35 sequence(TTGTTA) coresponded closely to the consensus sequences for Bacillus subtilis RNA polymerase with major sigma factor. The guanine-plus-cytosine content of the coding region of the xylA gene was 56mol% while that of the third position of the codons was 63 mol%. Based on the comparison with the amino acid sequences of several other carbohydrate degrading enzymes, two conserved regions, possibly participating in the catalytic mechamism of $\beta $-xylosidase xylA, were identified in 278-298 and 329-350 regions of the translated xylA gene. The nucleotide sequence of the xylA was found to exhibit no homology to any other genes so far reproted.