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dc.description.abstractRho protein, a member of small GTP-binding proteins, is involved in regulating the cytoskeletal structure in yeasts and mammalian cells. According to recent study, the function of rho in plants is also involved regulation of cytoskeleton, especially tip growth of pollen tube. In this study, a putative rho gene isolated from mung bean cDNA library, named as rho1Vr, was studied to understand its characters. To characterize of protein product of putative rho1Vr gene, the rho1Vr cDNA was overexpressed in E. coli. Its protein was purified, and is about 23 KDa size. The recombinant putative Rho1Vr protein have a specificity on guanosine nucleotide and a intrinsic GTP hydrolysis activity. The rate of intrinsic GTP hydrolysis was about 0.0045 mol/min/mol at 37 ℃, which is comparable with other small GTP-binding proteins. These facts suggest that the isolated putative recombinant Rho1Vr protein is a typical small GTP-binding protein. The promoter fragment of putative rho1Vr was isolated by IPCR. It contains TATA box for binding of RNA polymerase II and binding sites on two transcription factor, P and SBF-1, identified in plant.;Small GTP-binding protein 중에 하나인 Rho 단백질은 yeast와 포유동물 세포에서 세포내골격 구조와 관련된 세포 신호 전달 과정에 핵심적인 역할을 담당하고 있는 것으로 보고되고 있다. 최근 연구에 따르면, 식물에서도 rho 의 기능 또한 세포내골격, 특히 꽃가루관의 종단성장의 조절에 관련되어 있음이 알려져 있다. 녹두의 cDNA library로부터 분리된 rho 유전자인 rho1Vr은 그것의 특징과 기능을 이해하기 위해 조사된 결과 다음과 같은 결론을 얻었다. 1. 재조합된 Rho1Vr 단백질은 크기가 23 KDa이다. 이것은 rho 유전자에서 추론된 아미노산의 크기인 23 KDa과 그리고 Northern 혼성화 분석에서 알려진 rho1Vr 전사체의 크기인 1,100 개의 뉴클레오티드와 상응한다. 2. rho1Vr 유전자의 재조합 단백질 산물은 small GTP-binding protein이 갖는 공통된 특징인 구아노신 뉴클레오시드에 대한 특이성과 단백질 자체에서 갖는 GTP 분해 능력을 갖는다. 이것은 분리된 Rho1Vr 단백질이 전형적인 small GTP- binding protein임을 제안한다. 3. rhD1Vr유전자의 조절 부위의 일부분이 분리되었다. 그것은 749 뉴클레오티드로 이루어져 있고, TAT A box와 식물에서 알려진 두 가지의 조절물질에 대한 부착위치로 예상되는 서열을 포함한다.-
dc.description.tableofcontentsAbstract I. Introduction = 1 II. Materials and Methods 1. Materials (1) Experimental plants = 6 (2) General chemicals = 6 2. Methods (1) purification of rho1Vr protein products 1) Construction of rho1Vr overexpression plasmid = 6 2) Cultivation of the putative rho1Vr overexpression strain and purification of recombinant Rho1Vr proteins = 10 (2) Characterization of Rho1Vr recombinant proteins 1) Specificity of nucleotide binding = 11 2) Determination of GTP hydrolysis activity = 12 (3) Isolation of rho1Vr promoter fragment 1) Preparation of genomic DNA from mung bean = 13 2) Isolation of putative rho1Vr promoter fragment from genomic DNA by inverse PCR amplification = 14 3) Cloning of PCR product and sequence analysis = 14 III. Results & Discussion 1. Characterization of putative Rho1Vr recombinant proteins (1) Overexpression and purification of putative Rho1Vr proteins in E. coli = 16 (2) Characterization of putative Rho1Vr proteins = 18 2. Isolation and analysis of rho1Vr promoter fragment = 21 IV. Conclusions = 30 V. References = 31 Abstract in Korean = 36-
dc.format.extent1486243 bytes-
dc.publisher이화여자대학교 대학원-
dc.title녹두 rho 유전자의 발현 산물과 promoter에 관한 연구-
dc.typeMaster's Thesis-
dc.format.page37 p.-
dc.identifier.major대학원 생물과학과- 2-
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