Title Page

ABSTRACT

Contents

General Introduction 10

Chapter 1. DNA transposons: a review of the mechanisms underlying and the significance of protein expression techniques 11

1.1. Introduction 12

1.2. Characteristics of transposons involved in protein production 15

1.3. Transposon metabolism involved in protein production 18

1.4. Transposons as a strategy to increase protein production 21

1.5. Conclusions and Perspectives 26

Chapter 2. Improvement of Tol2 transposon system by modification of Tol2 transposase 27

2.1. Introduction 28

2.2. Materials and Methods 30

2.1. Cell culture 30

2.2. Plasmid design and construction 30

2.3. Transfections 30

2.4. Measurement of luciferase activity 30

2.5. Immunofluorescence 31

2.6. Statistical analyses 31

2.3. Results 32

2.3.1. Effect of NLS-H2B-Tol2 transposase on protein production 32

2.3.2. Identification of the mechanism by which NLS-H2B-Tol2 transposase enhances protein production 36

2.3.3. Importin beta maximizes the effect of NLS-H2B-Tol2 transposase on protein production 38

2.4. Discussion 42

2.5. Conclusions 43

References 44

국문초록 48

Figure 1.1.1. The categories of transposable elements are divided into various subcategories. 14

Figure 1.2.1. Sleeping Beauty (SB), piggyBac, Tol2 autonomic transplant element, transposase tissue and functional domains. 17

Figure 1.3.1. Transposition model based on cut-and-paste. 20

Figure 1.4.1. Technology for protein expression using Sleeping Beauty (SB), piggyBac, and Tol2. 25

Figure 2.3.1.1. Effect of NLS-H2B-Tol2 transposase on protein production. 34

Figure 2.3.1.2. Maintaining improved productivity of NLS-H2B-Tol2 transposase in single cell clones. 35

Figure 2.3.2.1. Mechanism through which NLS-H2B-Tol2 transposase enhances protein synthesis. 37

Figure 2.3.3.1. Importin beta maximizes the effect of NLS-H2B-Tol2 transposase on protein production. 40