Title Page

Contents

Abstract 11

Ⅰ. INTRODUCTION 13

Ⅱ. MATERIALS AND METHODS 18

1. Bacterial strains and culture conditions 18

2. Creation of Pseudomonas aeruginosa lysogen 18

3. Preparation of phage lysate 19

4. Phage Infection Assay 19

5. PCR of phage DNA replication 19

6. qPCR for phage DNA standard curve 20

7. Transposon mutagenesis 20

8. Creation of the ligD deletion mutant 21

9. Sequence analysis 21

Ⅲ. RESULTS 25

1. Temperate phages display limited infectivity, with the imprint of the infected host strain 25

2. The limited phage infectivity correlates with the phage genome synthesis in the infected strains 28

3. Plaque formation efficiency varies by phage-producing strains 31

4. Two mutant lysogens were identified to produce progeny phages with altered limited infectivity 33

5. Altered limited infectivity of the progeny phages is associated with the phage genome, not with the host genome 36

6. Two mutant lysogens affect the limited infectivity of phages. 41

7. The whole genome sequencing of the progeny phages revealed codon changes in ORF13, ORF27 and 8 structural proteins 43

Ⅳ. DISCUSSION 48

Ⅴ. CONCLUSION 50

REFERENCES 51

ABSTRACT IN KOREAN 55

Table 1. Bacterial strains in this study. 22

Table 2. Primers used in this study 23

Table 3. Summary of transposon insertion site. 35

Table 4. Summary of MP29* mutation location. 45

Fig. 1. Identification of differential infectivity of P. aeruginosa phage MP29 from four strains. 26

Fig. 2. Identification of differential infectivity of P. aeruginosa phage PP7 from four strains. 27

Fig. 3. Verification of correlation with the DNA synthesis of the infecting MP29 phage in PCR. 29

Fig. 4. Schematic representation of limited infectivity . 30

Fig. 5. Standard curve and Fraction of infectious virion of MP29. 32

Fig. 6. Schematic representation of transposon mutagenesis and phenotype of MP29PAK mutants.[이미지참조] 34

Fig. 7. Altered differential infectivity of MP29PAK mutants.[이미지참조] 38

Fig. 8. Assessment into the altered limited infectivity of MP29*. 39

Fig. 9. DNA synthesis and fraction of infectious virions of MP29*. 40

Fig. 10. Altered differential infectivity of MP22, and DMS3 by transposants. 42

Fig. 11. Verification of mutation location in MP29*. 44