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Sequencing of U18-treated yeast of different genetic background

ABSTRACT: Sequencing of U18-treated yeast of different genetic background

PROVIDER: PRJNA1209836 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR31983663_1.fastq.gz	Fastqsanger.gz
	SRR31983663_2.fastq.gz	Fastqsanger.gz
	SRR31983664_1.fastq.gz	Fastqsanger.gz
	SRR31983664_2.fastq.gz	Fastqsanger.gz
	SRR31983665_1.fastq.gz	Fastqsanger.gz

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Project description:The evolutionarily conserved Delta-Notch cell-cell signal-transduction pathway regulates the determination of various cell fates, which are important for the morphogenesis and development of numerous organs and tissues in many vertebrates and invertebrates. The Dll1 gene was mutated by homologous recombination in embryonic stem cells such that amino acids 2-116 were replaced with an in-frame fusion of the lacZ gene of E.coli. Mouse Dll1 genomic clones were isolated from a strain 129/SvPas library. The replacement vector was constructed by inserting a 4 kb ClaI/XhoI fragment, which encodes a part of the beta-galactosidase gene followed by the PGK-neo cassette, and a 2.6 kb XhoI/EcoRI fragment as the 3M-bM-^@M-^Y homologous region of the replacement vector into pKS+. This vector was linearized with SalI and ClaI, and a 4.1 kb SalI/NcoI as the 5M-bM-^@M-^Y homologous region plus an NcoI/ClaI fragment (800 bp) completing the b-galactosidase gene, were inserted, resulting in the complete replacement vector. This vector was linearized with SalI and electroporated in R1 embryonic stem cells. Correctly targeted clones were identified and verified by Southern blot analysis using external probes from the 3M-bM-^@M-^Y and 5M-bM-^@M-^Y region of the targeted area, and were injected into C57BL/6J embryo to obtain germline transmission. The two analysed Dll1 mutant mouse lines carry the same mutation but are on different genetic background: 1. Heterozygous F1-animals were backcrossed several times to the 129SV/J wild type; 2. Heterozygous F1-animals were outcrossed 11 generation to C3HeB/FeJ wild type Four organs (liver, spleen, thymus, brain) of two Dll1 mutant mouse lines on different genetic background carrying the same mutation were analysed by cDNA microarray technology. Experiment include 4-5 biological replicates for reference (wildtype) and mutant animals. Up to 4 technical replicates for each mutant mouse were performed. As reference pooled RNA was used. 50% of the chip hybridisations are dye sway experiments.

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Sequencing of U18-treated yeast of different genetic background

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