Development of a versatile expression system and its application to the study on the transcriptional regulation of SV40 T antigen 高効率発現系の開発およびSV40T抗原による転写調節解析への応用
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著者
書誌事項
- タイトル
-
Development of a versatile expression system and its application to the study on the transcriptional regulation of SV40 T antigen
- タイトル別名
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高効率発現系の開発およびSV40T抗原による転写調節解析への応用
- 著者名
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金, 東玩
- 著者別名
-
キム, ドンワン
- 学位授与大学
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東京大学
- 取得学位
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医学博士
- 学位授与番号
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甲第9157号
- 学位授与年月日
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1991-03-29
注記・抄録
博士論文
目次
- TABLE OF CONTENTS / p3 (0004.jp2)
- ABSTRACT / p7 (0008.jp2)
- Section1.Construction of pEF vector plasmids / p9 (0010.jp2)
- INTRODUCTION / p10 (0011.jp2)
- MATERIALS AND METHODS / p12 (0013.jp2)
- (a)Enzymes,linkers / p12 (0013.jp2)
- (b)Plasmid DNA sequencing / p12 (0013.jp2)
- RESULTS AND DISCUSSION / p13 (0014.jp2)
- (a)Structure of human EF-1α chromosomal gene and construction of the plasmids pEF-2 and pEF-3 / p13 (0014.jp2)
- (b)Construction of the plasmids pEF-220,pEF-223 and pEF-204 / p13 (0014.jp2)
- (c)Construction of the plasmid pEF-321 / p17 (0018.jp2)
- Section2.Efficiency of pEF vector plasmids and expression of eukaryotic genes encoding simian virus 40 T antigen or human CD4 / p21 (0022.jp2)
- INTRODUCTION / p22 (0023.jp2)
- MATERIALS AND METHODS / p24 (0025.jp2)
- (a)Cell cultures and transfection / p24 (0025.jp2)
- (b)CAT assay / p24 (0025.jp2)
- (c)Immunofluorescence assay / p27 (0028.jp2)
- (d)Northern blot analysis / p28 (0029.jp2)
- (e)Staining of cell colonies with Giemsa's solution / p28 (0029.jp2)
- RESULTS AND DISCUSSION / p30 (0031.jp2)
- (a)Construction of plasmids pEF220-CAT,pEF223-CAT,pEF204-CAT and pEF321-CAT / p30 (0031.jp2)
- (b)Measurement of EF-1α promoter activity of pEF-CAT plasmids and identification of the promoter region of EF-1α gene necessary for efficient gene expression / p30 (0031.jp2)
- (c)Comparison of the CAT acticvity of pEF-CAT plasmids with that of pSV2-CAT plasmid in various cell types / p33 (0034.jp2)
- (d)Efficiency of stable expression by the pEF321 p1asmid / p37 (0038.jp2)
- (e)Expression of the simian virus 40(SV40) T antigen by the plasmids pEF204 and pEF321 / p44 (0045.jp2)
- (f)Expression of the cDNA encoding human CD4(cluster of differentiation N4)by pEF321 plasmid / p50 (0051.jp2)
- (g)Conclusions / p50 (0051.jp2)
- Section3.Study on the transcription regulatory function of simian virus 40(SV40) T antigen on its early promoter / p54 (0055.jp2)
- INTRODUCTION / p55 (0056.jp2)
- MATERIALS AND METHODS / p59 (0060.jp2)
- (a)Structure of pSV2-CAT plasmid and construction of pSV8-16-CAT plasmid containing origin-defective SV40 early promoter / p59 (0060.jp2)
- (b)Construction of plasmids pSVP-CAT and pSV8-16P-CAT containing enhancer-defective SV40 early promoter / p60 (0061.jp2)
- (c)Construction of the plasmid pSVd-CAT containing the SV40 early promoter defective in the T antigen binding sites I and II / p60 (0061.jp2)
- RESULTS / p63 (0064.jp2)
- (a)Autoregulation of origin-defective SV40 early promoter by T antigen / p63 (0064.jp2)
- (b)Autoregulation of origin-containing SV40 early promoter by T antigen in the presence or absence of enhancer sequence / p66 (0067.jp2)
- (c)Transactivation of SV40 early promoter by T antigen / p67 (0068.jp2)
- DISCUSSION / p73 (0074.jp2)
- REFERENCES to section1 and section2 / p77 (0078.jp2)
- REFERENCES to section3 / p81 (0082.jp2)
- ACKNOWLEDGMENTS / p84 (0085.jp2)