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대표형(전거형, Authority) | 생물정보 | 이형(異形, Variant) | 소속 | 직위 | 직업 | 활동분야 | 주기 | 서지 | |
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title page
Contents
ABSTRACT 8
Introduction 10
Materials and methods 16
Cell culture and virus infection. 16
Expression plasmid. 16
Construction of the truncation mutants of the 84-kDa UL112-113 proteins. 18
Construction of the UL44 truncation mutants. 19
Yeast two-hybrid assay. 20
In vitro transcription/translation. 22
Synthesis and purification of GST fusion protein. 23
In vitro binding assay (GST pull-down assay). 24
Transient DNA transfection. 24
Coimmunoprecipitaion. 25
Antibodies. 26
Indirect immunofluorescence assay (IFA). 27
Immunoblot analysis. 27
Result 29
Interaction of the UL112-113 proteins with UL44 in the yeast two-hybrid assay. 29
The UL112-113 proteins are associated with UL44, but not with cellular PCNA, in HCMV-infected cells. 32
UL44 interacts with all of the four different UL112-113 proteins in vitro. 35
Domains of the 84-kDa form required for interaction with UL44. 39
Both the C-termina1 region and the central dimerization domain of UL44 are necessary for binding with the 84-kDa UL112-113 protein. 42
Recruitment of UL44 to the pre-replication foci by UL112-113 proteins requires both coexpression of the UL112-113 proteins and UL44 binding. 48
Discussion 53
REFERENCES 58
국문초록 66
Figure 1. Interaction of the four UL112-113 proteins with UL44 (PPF) in yeast two-hybrid assays. 30
Figure 2. Interaction of the UL112-113 with UL44 or PCNA in virus-infected cells. 33
Figure 3. Interaction of the UL112-113 proteins with UL44 in vitro. 37
Figure 4. Coimmunoprecipitation of truncation mutant forms of the 84-kDa UL112-113 protein with UL44. 40
Figure 5. Construction of expression plasmid containing the 84-kDa (SD) cDNA. 43
Figure 6. Coimmunoprecipitation of the 84-kDa UL112-113 protein and the UL44 truncation mutants. 46
Figure 7. Effects of the expressing four UL112-113 proteins on the recruitment of UL44 to the pre-replication foci. 50
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