Please use this identifier to cite or link to this item: http://scholarbank.nus.edu.sg/handle/10635/12871
Title: Investigation of germline excision of transgene by using the cre/loxp recombination system in zebrafish
Authors: LIU XINGJUN
Keywords: Zebrafish, Cre/loxP, ZP3, ovary, transgenic, germline
Issue Date: 23-Mar-2007
Source: LIU XINGJUN (2007-03-23). Investigation of germline excision of transgene by using the cre/loxp recombination system in zebrafish. ScholarBank@NUS Repository.
Abstract: To test the Cre/loxP recombination system for germline transgene excision in zebrafish, a zebrafish genomic DNA clone containing three tandem-repeated oocyte-specific zp3 genes was isolated and characterized. The 5a?? flanking region from one of the zp3 genes was used to develop stable zp3:gfp transgenic zebrafish lines and faithful oocyte-specific GFP expression was observed, indicating that an individual zp3 promoter harbors all necessary elements for oocyte-specific transcription. Subsequently, a stable zp3:cre transgenic line with oocyte-specific Cre expression was generated by co-injection of a Cre expression construct, pZP3-CRE and a loxP-flanked reporter gene construct, pKRT8-LGLR. Although effective Cre-loxP recombination was observed in the transient expression system with the two constructs, no chromosomal DNA excision was detected by crossing the zp3:cre transgenic line with a stable loxP transgenic line. Our data indicated that the efficiency of Cre-mediated recombination at the chromosomal level was quite low, if present in zebrafish.
URI: http://scholarbank.nus.edu.sg/handle/10635/12871
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