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귀리 원형질체에서 피토크롬 작용에 의한 이노시톨 인지질 대사
채쾌,표택물,박문환,조태주 ( Quae Chae,Taeck Yul Pyo,Moon Hwan Park,Tae Ju Cho ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.2
The stimulation of breakdown of phosphatidylinositol 4,5-bisphosphate (PIP₂) by the irradiation of red light (660 nm) was investigated by TLC/autoradiography after incorporation of ^(32)Pi into the oat protoplasts. The highest breakdown of PIP₂ was observed at 5 seconds of incubation after irradiation of red light for 3 seconds. When various wavelengths of light were irradiated, the highest breakdown of PIP₂ was observed at 660 nm which is the absorption maximum of phytochrome Pr form. Inositol 1,4,5-trisphosphate (IP₃) which is a metabolic product of PIP₂ was identified in oat cell by Dowex anion column chromatography, using [³H]-IP₃ as a standard compound. The level of IP₃ was substantially increased by irradiation of red light for 3 seconds. In addition, transient changes of the cytosolic free Ca^(2+) concentration by the red light were also monitored with the fluorescent Ca^(2+) indicator Quin 2 in the presence and the absence of extracellular Ca^(2+). The results suggest that signal transduction via IP₃ and Ca^(2+) may also be feasible in the phytochrome mediated cell responses.
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조태주,정기아,채쾌 ( Tae Ju Cho,Kee A Chung,Quae Chae ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.5
To investigate how light regulates plant gene experssion, we initiated a research program to isolate light-regulated soybean cab genes encoding chlorophyll a/b-binding proteins. For this purpose, a soybean cDNA library was constructed and screened. The inserts from three out of the nine soybean λ cab clones obtained were subcloned into pBluescribe^+, and designated pGAB3, pGAB7 and pGAB11. The restiction maps of pGAB3 and pGAB7 were found to be considerably different from each other. Further characterization of the two clones by dot blot hybridization and sequence analysis showed that the two soybean cab clones represent distinct cab genes in soybean. pGAB3, which is very homologous to the pea cab clone pAB96, has a full-length cDNA insert of 1.0 kb in size. pGAB7 has a 0.85 kb insert containing a truncated coding sequence. The 3` untranslated regions of the pGAB3 and pGAB7 are so divergent that any homologous regions can not be identified. pGAB11 was found to be very homologous to the pGAB3, and it remains to be determined whether the pGAB11 represents a distinct cab gene.
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Isolation and Characterization of Chlorophyll a/b Binding Protein Genes in Soybean
조태주,정기아,채쾌,Cho, Tae-Ju,Chung, Kee-A,Chae, Quae 생화학분자생물학회 1991 한국생화학회지 Vol.24 No.5
빛에 의한 식물체 유전자 발현의 조절기작을 이해하기 위하여, 대두에서 빛에 의해 그 발현이 조절되는 유전자를 클로닝하는 작업을 일차적으로 실시하였다. cab은 엽록체에 존재하는 chlorophyll 결함 단백질의 유전자로, 빛의 존재하에서 또는 암소에서 키운 대두 종묘에서 추출한 mRNA를 사용하여 Northern blot hybridization을 수행한 결과, 대두의 cab 유전자 발현이 빛에 의해 대폭 증진됨이 확인되었다. 이에 대두의 cab 유전자를 클로닝하기 위해 cDNA library를 제조한 다음 screening 작업을 실시하였다. 분리 동정된 9개 의 ${\lambda}$ cab cDNA clone중 두 개 clone의 insert를 subcloning하여 각각 pGAB3와 pGAB7이라 명명하였다. 이들 두 cDNA clone들은 각각 다른 cab gene에 대한 cDNA clone인 것으로 밝혀졌으며, 3' untranslated region에서의 염기서열이 매우 다른 것으로 나타났다. pGAB3은 31 nucleotide의 5' untranslated region을 포함한 full-length cDNA clone이며, 32개 아미노산으로 구성된 transit peptide를 갖고 있는 것으로 나타났다. To investigate how light regulates plant gene experssion, we initiated a research program to isolate light-regulated soybean cab genes encoding chlorophyll a/b-binding proteins. For this purpose, a soybean cDNA library was constructed and screened. The inserts from three out of the nine soybean ${\lambda}$ cab clones obtained were subcloned into pBluescribe+, and designated pGAB3, pGAB7 and pGAB11. The restiction maps of pGAB3 and pGAB7 were found to be considerably different from each other. Further characterization of the two clones by dot blot hybridization and sequence analysis showed that the two soybean cab clones represent distinct cab genes in soybean. pGAB3, which is very homologous to the pea cab clone pAB96, has a full-length cDNA insert of 1.0 kb in size. pGAB7 has a 0.85 kb insert containing a truncated coding sequence. The 3' untranslated regions of the pGAB3 and pGAB7 are so divergent that any homologous regions can not be identified. pGAB11 was found to be very homologous to the pGAB3, and it remains to be determined whether the pGAB11 represents a distinct cab gene.
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