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Mutagenesis of sepiapterin reductase by imprecise excision of the piggyBac transposon in Drosophila melanogaster
piggyBac transposon의 imprecise excision에 의한 초파리 세피압테린 환원효소 돌연변이체 제작

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Authors
김희종
Advisor
임정빈
Major
자연과학대학 생명과학부
Issue Date
2013-02
Publisher
서울대학교 대학원
Keywords
피기백 트랜스포존초파리돌연변이세피압테린 환원효소
Description
학위논문 (박사)-- 서울대학교 대학원 : 생명과학부, 2013. 2. 임정빈.
Abstract
Mutagenesis by transposon-mediated imprecise excision is the most extensively used technique for mutagenesis in Drosophila. Although P-element is the most widely used transposon in Drosophila to generate deletion mutants, it is limited by the insertion coldspots in the genome where P-elements are rarely found. The piggyBac transposon was developed as an alternative mutagenic vector for mutagenesis of non-P-element targeted genes in Drosophila because the piggyBac transposon can more randomly integrate into the genome. Previous studies suggested that the piggyBac transposon always excises precisely from the insertion site without initiating a deletion or leaving behind an additional footprint. This unique characteristic of the piggyBac transposon facilitates reversible gene-transfer in several studies, such as the generation of induced pluripotent stem (iPS) cells from fibroblasts. However, it also raised a potential limitation of its utility in generating deletion mutants in Drosophila. In this study, I report multiple imprecise excisions of the piggyBac transposon at the sepiapterin reductase (SR) locus in Drosophila. Through imprecise excision of the piggyBac transposon inserted in the 5′-UTR of the SR gene, I generated a hypomorphic mutant allele of the SR gene which showed markedly decreased levels of SR expression. My finding suggests that it is possible to generate deletion mutants by piggyBac transposon-mediated imprecise excision in Drosophila. However, it also suggests a limitation of piggyBac transposon-mediated reversible gene transfer for the generation of induced pluripotent stem (iPS) cells.
Language
English
URI
https://hdl.handle.net/10371/121356
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College of Natural Sciences (자연과학대학)Dept. of Biological Sciences (생명과학부)Theses (Ph.D. / Sc.D._생명과학부)
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