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Soybean의 Small GTP-binding Protein (sRab7)이 Endocytosis에 결함이 있는 Yeast ypt7 mutant 를 Complements
천충일,양효숙 숙명여자대학교 자연과학연구소 1997 자연과학논문집 Vol.- No.8
포유류의 Rab7에 대한 콩과식물의 homolog(sRab7)에 대한 cDNA가 soybean에서 분리된 바 있다. 이 단백질은 small GTP-binding protein에 특이한 공통 서열 motif(G1-G5)를 갖고 있으며, 세포내 수송신호로 알려진 C-terminal부위를 갖고 있다. sRab7이 정확한 Ypt7p(Rab7)의 homolog인지를 확인하기 위하여, srab7 cDNA가 YEp51의 GAL10 promoter의 downstream에 삽입되었다. 형질전환된 yeast mutant는 mutant와 대조적으로 하나의 큰 vacuole을 갖고 있음이 확인되었다. 이 발견은 sRab7이 yeast Ypt7p(Rab7)과 기능적으로 동일함을 보여준다. A cDNA encoding legume homolog of mammalian Rab7(sRab7) was isolated from soybean. sRab7 has the conserved wequence motifs (G1-G5) which are specific to small GTP-binding protein and C-terminal region known as intracellular targeting signal. In order to confirm that sRab7 is an authentic Ypt7p(Rab7) homolog, the srab7 cDNA was inserted downstream of the GAL10 promoter in YEp51(pHYE51). The transformed mutant showed a single large vacuole in contrast to mutant. Out finding suggests that sRab7 was confirmed to be a functional counterpart of yeast Ypt7p(Rab7).
Small GTP-binding Protein, Rab7 Homologs의 Molecular Cloning
양효숙,천충일 숙명여자대학교 자연과학연구소 1996 자연과학논문집 Vol.- No.7
Soybean의 뿌리혹에서 두개의 small GTP-binding protein(sRab1, sRab7)이 질소고정시 유도됨이 밝혀졌고, cDNA가 분리된 바 있다. rab7의 유도되는 정도가 높은 점에 착안해 cDNA library screening을 실시한 결과, 추가로 3개의 sRab7과 유사한 단백질들이 cloning되었다(sRab71, -b, 그리고 -c). 이들은 DNA sequence 상에서 서로 높은 homology를 보이며, srab7 gene과도 높은 homology를 갖는다. 뿐만 아니라, small GTPase가 갖는 conserved sequence motifs(G1 to G5)를 보이고, intracellular targeting signal에 관련된 것으로 알려진 C-terminal region에는 다른 Rab proteins과 매우 다른 amino acid sequences를 갖는다. 반면에 sRab7과 또한 각 homologs간에는 C-terminal region까지도 높은 homology를 보였다. 이러한 결과는 soybean nodules의 발생시, endocytosis에 필요한 small GTP-binding protein이 여러 homology 의 형태로 존재하며, 이들의 세분된 각각의 기능에 대한 연구가 필요함을 보여주고 있다. cDNA's of two small GTP-binding proteins (sRab1, sRab7) were isolated and shown to be induced during nitrogen fixation. Since the rab7 induction was very high, additional screening cDNA library of soybean resulted in isolation of four more sRab7 homolotys (sRab7a, -b and -c). They have high homology to each other at DNA levels, and to srab7, too. They also show the conserved sequence motif (G1 to G5) which are specific to small GTPase. These clones have characteristic C-terminal region known as intracellular targeting signal which are highly homologous to sRab7 although sRab7d shows high homology to Rab7. We conclude that during soybean nodulation small GTP-binding proteins required for endocytosis may exist as many homologs which need to be studied in detail for the function of each homologs.
오혜숙,천충일 숙명여자대학교 자연과학연구소 2000 자연과학논문집 Vol.- No.11
식물은 water stress를 Abscisic Acid (ABA)에 의한 신호전달 과정에 의해 반응하는 것으로 알려져 있는 데, homeobox gone인 Athb-12는 ABA에 의해 발현이 유도되는 것이 보고되어 water stress에 중요한 역할을 할 것으로 예상되었다. Athb-12유전자의 promoter지역 약 2.1 Kb를 β-glucuronidase (GUS)와 fusion하여 transgenic tobacco를 제작하였다. 이를 이용하여 ABA처리시 어떠한 양상으로 발현하는 지를 관찰하였다. 또한 다른 환경stress에 어떻게 반응하는지 조사하기 위해 UV처리와 wounding처리를 하여 GUS의 발현을 관찰하였다. 이 결과들은 Athb-12가 ABA에 의한 반응 뿐 아니라, 전반적인 스트레스들의 반응에 관여할 가능성을 보여 주었다. It has been known that plant responds to water stress by ABA-mediated signal transduction. Since Athb-12, a homeobox gene from Arabidopsis, was reported to be inducible to ABA, it was expected to be important to plant response to water stress. About 2.1 kb DNA of Athb-12 promoter region was fused to f-glucuronidase gene and the resulting construct was used in making transgenic tobacco plant. ABA-treatment to the transgenics was observed. Other environmental stresses such as UV and wounding were also tested. The results obtained showed a possibility that Athb-12 is involved in plant responses to most environmental stresses as well as ABA.
Methylmethacrylate와 n-Butylalcohol의 에스테르 교환반응에 관한 연구
鄭舜旭,朴根浩,孫秉淸,金忠一 弘益大學校 1988 弘大論叢 Vol.20 No.2
The transesterification reaction between methylmethacrylate and n-butyl alcohol was kinetically investigated in the presence of constant quantity of inhibitor and various metal acetate catalyst at 120℃. The quantity of n-butylmethacrylate produced in the reaction flask was measured by gas-chromatography, and the reaction rate was investigated by measuring of the quantity of the product the reaction under various catalysts. The transesterification was carried out in the first order reaction kinetics with respect to the concentration of methylmethacrylate and n-buty1 alcohol, respectively. The linear relationship was show between rate constant and absolute temperatures, and the activation energy of 14.7Kcal for the reaction with zinc acetate catalyst was calculated. The maximum reaction rate was appeared at the range of 1.5 to 1.6 of electro-negativity of metal ions.
애기장대의 Athb-12 유전자에 대한 불활성 돌연변이의 선발
손오라,이현정,천충일 숙명여자대학교 자연과학연구소 2001 자연과학논문집 Vol.- No.12
애기장대의 homeobox gene인 Athb-12는 ABA에 의해 발현이 유도되는 것이 보고되어 water stress에 중요한 역할을 할 것으로 예상되었다. 이 유전자의 기능을 연구하기 위해, knockout mutants를 찾고자 하였다. T-DNA insertion에 의한 불활성화 돌연변이들의 pools을 이용하여 polymerase chain reaction(PCR)에 의해 pools을 screen하였다. 1차 screen에서 정확하게 Athb-12유전자에 insertion된 돌연변이체를 Southern blots 및 DNA sequencing으로 확인하였고, 이들을 2차 screen에서도 성공적으로 맞는 mutants를 선발하였다. 최종적으로 mutant를 찾아내면 Athb-12의 기능연구에 유용하리라 예상된다. Sine the Athb-12 gene, a homeobox gene from Arabidopsis, was reported to be expressed by ABA induction, it was expected to be important in plant response mechanism to water stress. Screening for knockout mutant for the gene was performed to study the function of the gene. Polymerase Chain Reactions(PCR) were used for screening pools of T-DNA inserted inactivated mutants. From the primary screening, the exact inserted mutant at the Athb-12 gene was found and confirmed by Southern blots and DNA sequencing. The correct mutant was successfully selected from the secondary screening, too. Finding the final insertion mutant will be useful in studying the function of Athb-12.