Abstract
BackgroundTransgenic gpt delta mouse and rat models were developed to perform gpt and Spi− assays for in vivo mutagenicity tests. The animals were established by integration of lambda EG10 phage DNA as a transgene into the genome. The inserted position of the transgene on chromosome was determined by fluorescent in situ hybridization and Southern blot analyses; however, the exact position and sequence of the inserted junction were not known. To identify the site and pattern of genomic integration of the transgene copies, genomic DNAs extracted from C57BL/6J gpt delta mice and F344 gpt delta rats were applied to whole genome sequencing and mate-pair analysis.ResultsThe result confirmed that multi-copy lambda EG10 transgenes are inserted at a single position in the mouse chromosome 17. The junction contains 70 bp of overlapped genomic sequences, and it has short homology at both ends. A copy number analysis suggested that the inserted transgenes may contain 41 head-to-tail junctions and 16 junctions of other types such as rearranged abnormal junctions. It suggested that the number of intact copies could be approximately 40 at maximum. In the F344 gpt delta rats, transgenes are inserted at a single position in the rat chromosome 4. The junction contains no overlapped sequence but 72-kb genomic sequence including one gene was deleted. The inserted transgenes may contain 15 head-to-tail junctions and two rearranged junctions. It suggested that the number of intact copies could be 14 at maximum. One germline base substitution in the gpt gene rescued from gpt delta rats was characterized.ConclusionsThe exact inserted positions of the lambda EG10 transgene in the genome of gpt delta transgenic rodents were identified. The copy number and arrangement of the transgene were analyzed. PCR primers for quick genotyping of gpt delta mice and rats have been designed.Electronic supplementary materialThe online version of this article (doi:10.1186/s41021-015-0024-6) contains supplementary material, which is available to authorized users.
Highlights
Transgenic gpt delta mouse and rat models were developed to perform gpt and Spi− assays for in vivo mutagenicity tests
Lambda EG10 transgenes are inserted at this position
Genomic integration of the lambda EG10 transgene of gpt delta transgenic rodents was analyzed by whole genome sequencing and mate pairs (MPs) analysis
Summary
Transgenic gpt delta mouse and rat models were developed to perform gpt and Spi− assays for in vivo mutagenicity tests. The animals were established by integration of lambda EG10 phage DNA as a transgene into the genome. Transgenic rodent gene mutation assays are useful tools to detect in vivo mutagenicity in various types of rodent tissues [1]. These assays are based on transgenic animals that contain multiple copies of chromosomally integrated shuttle vectors that harbor reporter genes for the detection of gene mutations. The transgenic mouse gpt delta was established via the microinjection of lambda EG10 phage shuttle vectors into the fertilized eggs of C57BL/6J mice [8]. Lambda EG10 DNA is 48 kb in size, and multiple copies of the transgene are integrated in a single
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
