第一个书签之前
由正交融合蛋白激活的酿酒酵母合成启动子库的构建
Comstruction Of A Library Of Yeast S. cerevisiae Synthetic Promoters Activated By Orthogonal Chimeric Proteins
摘要
Abstract
Contents
Chapter 1 Introduction
1.1 Background and significance of this study
1.2 Biosensors that respond to β-estradiol
1.3 Synthetic promoter design with lexOp2 operators
1.4 The basic parts used in this project
1.5 Research status in this direction
1.6 Main research content and goal
1.6.1 Testing the biosensor performance with various distance betweenlexOp2 operators
1.6.2 Testing the biosensor performance with various distance betweenlexOp2 operator and TATA box
Chapter 2 Empirical methods
2.1 Laboratory machine
2.2 Experimental materials
2.2.1 Microbiology Strain
2.2.3 Enzymes
2.3 Protocols
2.3.1 Yeast genome extraction methods
2.3.2 Bacteria transformation
2.3.3 Restriction digest
2.3.4 Gel electrophoresis
2.3.5 DNA elution
2.3.6 Primer preparation
2.3.7 Blackburn PCR
2.3.8 Isothermal assembly—Gibson protocol
2.3.9 Touchdown PCR
2.3.10 Making Competent Cells
2.3.11 Mini preparation
2.3.12 Yeast transformation
2.3.13 Cryostocks
2.3.14 Colony picking
2.3.15 Yeast cell induction
2.2.16 FACS measurements—BD FACSCalibur
2.2.17 β -estradiol concentration curve preparation
2.2.18 Data analysis program
Chapter 3 Optimization based on various promotersand activation domain
3.1 Performance based on various promoter in the reportersection
3.2 Performance based on various promoters in the sensor section
3.3 Performance based on various activation domains in thesensor section
3.4 Chapter brief summary
4.1 Synthetic activated promoters in the literature
4.2 Performance based on various design of lexOp2 operators inthe reporter section
4.3 Performance based on an increasing number of lexOp2operators
4.4 Chapter brief summary
Chapter 5 Testing the biosensor performance withvarious positions of the lexOp2 operators
5.1 Biosensors based on different distance between two adjacentlexOp2 operators
5.1.1 Construction of biosensors with various distance between lexOp2operators
5.1.2 Performance of the biosensor with various distance betweenlexAop2 operators
5.1.3 Promoter comparison
5.2 Biosensors based on different distance between a lexOp2operator and the TATA box
5.2.1 Construction of biosensors with various distance between alexOp2 operator and the TATA box
5.2.2 Performance of the biosensors with various distance between asingle lexOp2 operator and the TATA box
5.2.3 Promoter comparison
5.3 Chapter brief summary
References
哈尔滨工业大学学位论文原创性声明和使用权限
Acknowledgements
