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퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석
차준영,네티엘마와티,김순길,이증주,임채오,정우식,이곤호,손대영,Cha, Joon-Yung,Netty Ermawati,Kim, Soon-Gil,Lee, Jeung-Joo,Lim, Chae-Oh,Chung, Woo-Sik,Lee, Kon-Ho,Son, Dae-Young 한국식물생명공학회 2003 식물생명공학회지 Vol.30 No.4
토양 내의 고농도의 염은 심각한 환경스트레스 중의 하나로 농작물의 생산을 감소시킨다. 식물은 염 스트레스로부터 벗어나기 위하여 많은 단백질을 합성한다든지 유전자들의 발현을 조절하는 등 여러 가지 생리, 생화학적인 변화를 일으킨다. 퉁퉁마디는 우리나라에 자생하는 염생식물로 갯벌과 염전주위에서 생육한다. 퉁퉁마디의 생화학적, 분자생물학적 내염성 기구를 이해하기 위하여 differential display방법으로 NaCl에 의하여 발현이 증가되는 cDNA들을 분리하였다. 본 연구에서는 그 중 하나인 ShADL의 특성을 조사하였다. ShADL은fructose-1, 6-bisphosphate aldolase와 높은 유사성을 보였다. 이 유전자는 1293bp길이에 359개의 아미노산으로 구성된 open reading frame을 포함하고 있으며, 이로부터 추정되는 분자량은 39 kDa이었다. ShADL단백질은 애기장대의 aldolase와 86%의 높은 유사성을 나타내었으며 같은 염생식물인 com-mon ice plant의 adolase와는 78%의 유사성을 보였다. Northern 분석결과, ShADL 유전자는 NaCl의 농도가 증가함에 따라 발현량이 급격히 증가하는 것으로 나타났다. Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marshes and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display. Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCl concentrations.
김선호,이경희,김경은,정미순,임채오,이신우,정우식,Kim, Sun-Ho,Lee, Kyung-Hee,Kim, Kyung-Eun,Jung, Mi-Soon,Lim, Chae-Oh,Lee, Shin-Woo,Chung, Woo-Sik Korean Society of Life Science 2007 생명과학회지 Vol.17 No.9
칼모둘린은 칼슘과 결합하는 센서로써 다양한 칼모둘린 결합 단백질들과의 상호 작용을 통하여 세포 내에서 여러가지 기능을 조절한다. 진핵 생물들은 많은 종류의 칼모둘린 결합 단백질을 가지고 있기 때문에 이러한 단백질들의 분리와 특성 규명이 중요하다. 이미 여러 가지 방법들을 이용하여 칼모둘린 결합 단백질들이 분리되었고 이미 알려진 단백질의 구조적인 유사성을 토대로 더 많은 단백질들이 예측되었다. 우리는 애기장대에서 칼모둘린 결합 단백질의 분리와 특성 규명을 위해 형광 단백질과 융합된 칼모둘린 과발현 형질 전환체를 제조하여 공촛점 현미경과 Western blot 을 이용하여 과발현 형질 전환체를 선별하였다. 또한 형질 전환체 내의 칼모둘린이 칼모둘린 결합 단백질과 상호 작용함을 pull-down 분석을 통해서 확인하였다. 이러한 결과들을 토대로 칼모둘린 과발현 형질 전환체를 이용하여, 칼모둘린과 상호 작용하는 여러 가지 칼모둘린 결합 단백질들을 분리할 수 있을 것으로 기대된다. Calmodulin (CaM), a ubiquitous calcium-binding protein, regulates diverse cellular functions by modulating the activity of a variety CaM-binding proteins (CaMBPs). Because eukaryotes have multiple CaMBPs, it is important to isolate and characterize them in different tissues and conditions. So far a number of CaMBPs have been identified through classical screening methods. Many classes of proteins have been predicted to bind CaMs based on their structural homology with already known targets. In an effort to develop a method for large-scale analysis of CaMBPs in Arabidopsis, we have generated a transgenic plants overexpressing AtCaM2-GFP. We performed protein pull-down assay to test whether exogenously expressed AtCaM2-GFP proteins can interact with CaMBPs. The exogenously expressed AtCaM2-GFP could strongly interact with a CaMBP, AS1 protein. This result suggests that AtCaM2-GFP in transgenic plants may interact with many CaMBPs in plant cell. Therefore, we will be able to isolate kinds of CaMBPs by using these transgenic plants in many different tissue and environments.
전통발효식품으로부터 Chitin 분해 미생물의 분리 및 특성 규명
고보경,최인순,이상현,임채오,이성호,갈상완,최영주 Plant molecular biology and biotechnology research 2004 Plant molecular biology and biotechnology research Vol.2004 No.-
A bacterial strain CJ-3 which produced chitinase was isolated from Korean traditional soy sauce. Using 16S rDNA analysis, the strain CJ-3 was identified as Bacillus atrophaeus. The approximate molecular weight of the putative chitinase enzyme was 31.0 kDa and the enzyme activity was remarkably induced by addition of colloidal chitin (0.5, 1.0, 2.0%). The antioxidant activity was increased 53% by the browning reaction products of B. atrophaeus CJ-3. Escherichia. coli lipopolysaccharides (LPS)-induced production of nitric oxide (NO) was reduced up to 45% by the browning reaction product in RAW264.7 macrophage. Inhibition of cell viability in the presence of LPS was recovered to normal level by the browning reaction product. These results suggest that browning reaction of B. atrophaeus CJ-3 plays an important role for activation of immune system. B. atrophaeus CJ-3 exhibited optimum temperature and pH of 37℃ and pH 7.8~8.0, respectively. The major intracelluar free amino acid was determined to be glutamate.
퉁퉁마디로부터 염에 의하여 유도되는 Aldolase 유전자의 분리 및 발현분석
차준영,네티 엘마와티,김순길,이증주,임채오,정우식,이곤호,손대영 Plant molecular biology and biotechnology research 2003 Plant molecular biology and biotechnology research Vol.2003 No.-
Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marches and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display.Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86% identity with Arabidopsis and 78% with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCI concentrations.
Screening for Highly Susceptible Soybean Genotypes to Agrobacterium tumefaciens
LEE, SUNG HO,LIM, CHAE HO,CHUNG, GYU HWA,WON, JONG LAC,PARK, JOONG SUK,KIM, ZHOO HYEON 경상대학교 유전공학연구소 1989 遺傳工學硏究所報 Vol.8 No.-
This experiment was conducted to screen the highly susceptible soybean genotypes to Agrobacterium tumefaciens. Plants from 703 lines and 20 cultivars were inoculated with Agrobacteriun tumefaciens Nopaline type strain c58 and Octopine type strain 15955. About 97% of tested lines were infected by Agrobacteriun tumefaciens strain c58, which only two lines, B678 and B679 responded to Agrobacteriun tumefaciens strain 15955. The number of highly susceptible lines with higher than 50% tumorigenesis and greater than 1 mm tumor size was 53, while 20 lines were not infected at all. Among the highly susceptible group, the lines 53, 79, 463, 551, 674, 675, 677, 678, and 679 could be used for transformation studies due to their high tuomrigenesis and large tumor size. The lines selected in the field test according to the degree of susceptibility showed the same responses to Agrobacterium infection in vitro.