In vivo excision and amplification of large human genomic segments using Cre/loxP- and large T antigen/SV40 ori-mediated machinery = Cre/loxP-, large T antigen/SV40 ori-시스템을 이용한 인간유전체 절편의 인체세포 내에서의 분리와 증폭

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In vivo excision and amplification of pre-determined, large genomic segments, directly from the genome of a natural host could be a novel cloning method with applications such as gap sequencing, gene therapy, and functional analysis of unknown genes in human cells. In this study, an in vivo excision and amplification system for human cells was devised with the Cre/loxP system of bacteriophage P1 and large T antigen/SV40 ori system of Simian virus 40. Trans-acting gene cre and large T antigen, each of which was under the control of a tetracycline responsive promoter, were inserted into the 5`- and 3`-UTR regions of the iNOS gene, respectively, by homologous recombination. Two loxP sequences, each of which serves as the recognition site for recombinase Cre, were also integrated unidirectionally into the 5`- and 3`-UTR regions of the iNOS gene. SV40 ori sequence, which serves as a conditional replication system, was inserted between the loxP sequences. The human cell line carrying the inserted elements was stably maintained until the excision and amplification functions are triggered by the induction of Cre and large T antigen, respectively. Upon induction by doxycycline, the 50-kb iNOS genomic region of human chromosome 17 flanked by two loxP sites was excised successfully.
Advisors
Kim, Sun-Changresearcher김선창researcher
Description
한국과학기술원 : 생물과학과,
Publisher
한국과학기술원
Issue Date
2001
Identifier
165898/325007 / 000993572
Language
eng
Description

학위논문(석사) - 한국과학기술원 : 생물과학과, 2001.2, [ iii, 34 p. ]

Keywords

in vivo excision and amplification; large T antigen/SV40 ori system; 인간유전체 절편의 분리와 증폭; Cre/loxP-system

URI
http://hdl.handle.net/10203/27938
Link
http://library.kaist.ac.kr/search/detail/view.do?bibCtrlNo=165898&flag=dissertation
Appears in Collection
BS-Theses_Master(석사논문)
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