식물 Polyol 들의 분리 및 대사

박훤범,윤정원 수원대학교 산업기술연구소 2002 산업기술연구소논문집 Vol.17 No.-

In the korean soybean seedlings O-methyltranferase and ononitol epimerase lead to accumulation of methylated inositols, D-ononiton and D-pinitol, while serve as osmorprotectants. This paper describes analysis for myo-inositol, D-ononitol, D-pinitol and glucose by TLC for metabolites in salt stressed soybean seedlings. The crude enzymes, myo-inosito-6-O-methyltranferase and ononitol epimerase, extract from soybean seedlings, which analysis by DEAE-cellulose column. This enzymes react with glucose to produce polyol metabolites.

대두에서 phyA 유전자의 promoter 분리

박훤범 수원대학교 자연과학연구소 2001 자연과학논문집 Vol.4 No.-

The promoter of phyA gene has been cloned by screening of genomic DNA library of soybean using a phytochrome cDNA as a probe. BlastN search has revealed several cis-acting DNA element in the promoter of phyA gene.

벼 호분충에서 Gibberellin 신호전달에 관여하는 Phosphatase 2C

박훤범 수원대학교 기능성생명소재연구소 2005 자연과학연구논문집 Vol.4 No.1

Limited information is available about the phosphorylation and dephosphorylation of many plant hormone-signaling proteins. We have identified one GA-inducible phosphatase 2C in rice. To check the function of this phosphatase 2C, the T- DNA insertional mutant was isolated and characterized. The phenotype of this phosphatase 2C mutant is quite different with the 10 days old seedlings of wild type. Their height is shorter than the wild type, and they show reduced lateral root formation. The phenotype of this mutant is co-segregated with the T-DNA. The induction of GA-inducible α-amylase in the aleurone is delayed in this T-DNA insertional mutant comparing with the wild type. By using anti-ⓟ-Thr and anti-ⓟ-Ser antibodies. the substrate of this phosphatase 2C was identified. A 65 kDa protein is phosphorylated in this T-DNA insertional mutant, but it is dephosphorylated in the wild type during early germination stage. To further characterize this 65 kDa protein, 2D gel electrophoresis and MALDI-TOF MS analysis is on progress.

대두에서 엽록체 ABC 단백질 유전자의 분리 및 연구

박훤범 수원대학교 기능성생명소재연구소 2003 자연과학연구논문집 Vol.2 No.2

In the course of a study concerning the molecular mechanisms of hypocotyl elongation during soybean seedlings which were grown in darkness, we generated a lot of ESTs from a cDNA library prepared from the dark grown soybean seedlings. One of the ESTs has a high similarity with a Arabidopsis thailana plastidic ATP-binding-cassette (atABC) protein in amino acid level. The putative soybean ABC protein contains an N-terminal transit peptide which targets it into chloroplast same as atABC1 protein. The transcription level of putative soybean ABC gene has investigated under the continuous red light, continuous far-red light, and complete dark condition. The main function of atABC1 protein is the transport of protoporphyrinogen Ⅸ which is the precursor of chlorophyll from the cytoplasm to the chloroplast.

메밀에서 비생물학적 스트레스에 의한 D-chiro-inositol의 함량 증가

박훤범 수원대학교 기능성생명소재연구소 2007 자연과학연구논문집 Vol.6 No.1

D-chiro-inositol which is the isomer of myo-inositol is well known drug for the type Ⅱ diabetes. The methylated form of D-chiro-inositol, pinitol and D-chiro-inositol are synthesized when the plants are exposed to the abiotic stresses such as drought, salinity and low temperature as osmoprotectants. In soybean, myo-inositol is converted to ononitol by O-methyltransferase and ononitol is converted to pinitol by ononitol epimerase and finally converted to D-chiro-inositol by demethylase. However there have been some reports that in buckwheat, myo-inositol tan be converted to D-chiro-inositol directly Thus we checked the cyclitol contents after abiotic stresses in buckwheat, and confirmed that myo-inositol is directly converted to D-chiro-inositol by myo-inositol epimerase.

대두 엽록체 ABC transporter 유전자의 애기장대에서의 대량발현으로 인한 엽록소 함량의 증가

박훤범 수원대학교 기능성생명소재연구소 2006 자연과학연구논문집 Vol.5 No.1

The soybean ABC protein contains an N-terminal transit peptide which targets it into chloroplast. To confirm the subcellular localization of soybean ABC protein experimentally, the transit peptide of soybean ABC transporter was fused to GFP protein. After transformation into the Arabidopsis protoplast, the localization of the GFP fusion protein was examined by confocal microscopy. It was confirmed that localization of the soybean ABC protein is chloroplast. The over-expression of soybean ABC transporter gene into Arabidopsis increases the chlorophyll contents in transgenic plant because the main function of soybean ABC protein is the transport of protoporphyrin IX which is the precursor of chlorophyll from the cytoplasm to the stroma of chloroplast.

대두에서 ononitol epimerase를 coding 하는 유전자의 분리와 E. coli에서 발현 및 기질 특이성 확인

박훤범 수원대학교 기능성생명소재연구소 2004 자연과학연구논문집 Vol.3 No.2

Myo-inositol plays a key role in plant development and growth. Conversion of glucose-6-phosphate to myo-inositol-1-phosphate is the first committed step in myo-inositol biosynthesis. Methylation and isomerization of myo-inositol forms 0-methyl inositols (ononitol, pinitol, etc.) which participate in abiotic stress related reponses, storage of seed products, and production of inositol-glycosides. Upon salinity stress, an 0-methyl transferase methylates C4 of myo-inositol then produces ononitol. The next step is epimerization of Cl of ononitol by ononitol epimerase then produces pinitol. However, there has been no report about the ononitol epimerase in plant yet. We have searched salinity inducible ESTs which were homologous with the NAD-dependent epimerase from the soybean EST data base. We have chosen one putative ononitol epimerase EST and sequenced the full-length cDNA. The putative ononitol epimerase cDNA was cloned into E. coli expression vector pET15b and we have checked the substrate of this putative ononitol epimerase, and found out that the substrate was ononitol by GC-FID. The transcripts level of the ononitol epimerase gene under the salinity condition was also checked.

Arabidopsis Genome Database 검색을 통한 추정 G-protein γsubunit 유전자의 동정

박훤범 수원대학교 자연과학연구소 1998 자연과학논문집 Vol.1 No.-

Heterotrimeric G-proteins relay signals from various seven trans-membrane domain receptors to intracellular effectors. Genes for G-protein α subunit and β subunit have been cloned in Arabidopsis, but a gene for G-protein γ has not been cloned yet. All the G-protein γ subunits so far cloned have a notable homologous region which is the CAAX sequence found at the carboxyl terminus of the known γ subunits, where A is an aliphatic amino acid and X is any amino acid. Arabidopsis Genome Database searches were performed and we found 10 non-redundant proteins which have CAAX sepuence at the carboxyl terminus. Among 10 proteins, one is very promising in terms of molecular weight, secondary structure prediction, and it has a prenyl group-binding site.

애기장대에서 GmNAP1의 과발현으로 인한 엽록소 함량 증가

박훤범(Phun Bum Park),안철현(Chul-Hyun Ahn) 한국생명과학회 2010 생명과학회지 Vol.20 No.10

암(dark) 상태에서 재배한 대두의 하배축 길이 생장의 분자 기작을 연구하기 위한 일환으로 암 상태에서 재배한 대두 하배축으로부터 cDNA library를 제작한 후 ESTs를 구축하였다. 이들 ESTs 중 색소체 ABC 단백질과 아미노산 서열이 매우 유사한 clone을 선발한 후 이 유전자의 전체염기서열을 결정하였다. GmNAP1 단백질은 엽록체로 향하는 transit peptide 서열이 존재한다. 빛에 의해 GmNAP1 유전자 전사가 어떻게 변화되는지 알아보기 위해 지속적인 적색광, 근적색광 그리고 암 상태에서 성장시키면서 유전자의 전사량을 확인하였다. 이 색소체 NAP1는 엽록소의 전구 물질인 protoporphytin IX를 세포질에서 엽록체로 이동시키는 기능을 한다. 대두에서 분리된 GmNAP1 유전자의 기능을 확인하기 위하여 35S 프로모터 뒤에 GmNAP1 유전자를 접합한 후 애기장대에 형질전환하였다. 형질전환 된 애기장대의 엽록소 함량은 야생형의 엽록소 함량보다 훨씬 높게 측정되었다. In the course of a research concerning the molecular mechanism of hypocotyl elongation that occurs during soybean seedling growth in darkness, we have generated a number of ESTs from a cDNA library prepared from the hypocotyls of dark-grown soybean seedlings. Comparison of the ESTs assigned a cDNA clone as a putative plastidic ATP-binding-cassette (ABC) protein homologue. The soybean GmNAP1 protein contains an N-terminal transit peptide which targets it into the chloroplast. The transcription level of the GmNAP1 gene was investigated under continuous red light, continuous far-red light, and complete darkness. The main function of this NAP1 protein is the transport of protoporphyrin IX which is the precursor of chlorophyll from the cytoplasm to the chloroplast. The GmNAP1 gene was transferred into the Arabidopsis under the CaMV 35S promoter. The chlorophyll level of this transgenic Arabidopsis plant was much higher than the chlorophyll level of the wild type Arabidopsis plant.

Increased Abiotic Stress Tolerance by Over-expressing OsABF2 in Transgenic Arabidopsis thaliana

Phun Bum Park(박훤범) 한국생명과학회 2012 생명과학회지 Vol.22 No.11

식물호르몬인 abscisic acid (ABA)는 식물의 비생물학적 스트레스의 적응과정에서 중요한 역할을 수행하고 있다. 또한 ABA는 종자휴면, 발아, 세포분열의 저해, 기공개폐와 같은 중요한 과정에 관여하고 있다. OsABF2 (Oryza sativa ABRE Binding Factor2)는 벼에서 비생물학적 스트레스와 ABA 신호전달 과정에 양성적으로 관여하는 bZIP 형태의 전사인자이다. OsABF2 유전자의 발현은 ABA와 다양한 스트레스 처리에 의해 유도된다. 본 논문에서는 OsABF2 유전자를 과발현한 애기장대가 가뭄, 고염, 고온 상태에서의 생존율이 야생형보다 증가하는 것을 확인하였다. 또한 ABA가 존재하는 상황에서 OsABF2 유전자를 과발현한 애기장대의 발아율이 감소하는 것을 확인하였다. 이러한 결과로 미루어 OsABF2 유전자를 과발현한 애기장대는 비생물학적 스트레스에 대한 내성이 증가하고 ABA 감수성은 증가하는 것으로 확인되었다. The phytohormone abscisic acid (ABA) plays an important role in the adaptive response of plants to abiotic stresses. ABA also regulates many important processes, including seed dormancy, germination, inhibition of cell division, and stomatal closure. OsABF2 (Oryza sativa ABRE binding factor2) is one of the bZIP type transcription factors, which are involved in abiotic stress response and ABA signaling in rice. Expression of OsABF2 is induced by ABA and various stress treatments. Findings show that survival rates of OsABF2 over-expressing Arabidopsis lines were increased under drought, salt, and heat stress conditions. The germination ratio of OsABF2 over-expressing Arabidopsis lines was decreased in the presence of ABA. Results indicate that OsABF2 over-expressing Arabidopsis lines have enhanced abiotic stress tolerance and have increased ABA sensitivity.