B6.FVB(Cg)-Tg(Colgalt2-EGFP/Rpl10a)DU9Htz/J

Stock number: 030257
Product name: Glt25d2-EGFP/L10a
Research areas: Neurobiology Research, Research Tools

Description

This bacTRAP transgenic strain, incorporating an EGFP-L10a ribosomal fusion protein targeted to the Colgalt2 gene, enables cell labeling, sorting, and affinity purification of cell-specific polysomal RNAs.

Detailed description

This bacTRAP line incorporates an EGFP-L10a ribosomal fusion protein targeted to the translational start site of the Colgalt2 (collagen beta(1-O)galactosyltransferase 2; also called Glt25d2) gene. The fusion protein enables affinity purification of CNS cell-specific polysomal RNAs which can be used for gene expression profiling.

Expression was carefully assayed by immunohistochemistry (IHC) with an antibody against enhanced green fluorescent protein (EGFP). The cerebellum, spinal cord, basal forebrain and corpus striatum, brainstem, and cerebral cortex were surveyed. Primary labeling was found to occur in layer 5b corticospinal, corticopontine pyramidal neurons, and small pyramidal cells. Fluorescence can also be visualized directly.

Images showing transgene expression in brain may be found on the Gensat gene expression and Gensat TRAP cell Showcase pages.

Development

A shuttle vector was used to introduce sequences encoding an EGFP-L10a ribosomal fusion protein into the translation start site of the Colgalt2 gene contained within BAC RP23-160M1. The modified BAC was introduced to FVB embryos. This strain was maintained on a mixed C57BL/6J, FVB, and Swiss Webster genetic background prior to being backcrossed to C57BL/6J for 9 generations by the donating laboratory. This is founder line DU9.

Allele

Symbol: Tg(Colgalt2-EGFP/Rpl10a)DU9Htz
Name: transgene insertion DU9, Nathaniel Heintz
MGI accession ID: MGI:5496687
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Tg(Colgalt2-EGFP/Rpl10a)DU9Htz
Marker name: transgene insertion DU9, Nathaniel Heintz
Chromosome: UN
Strain of origin: FVB/N
Expressed genes: GFP,Green Fluorescent Protein,; Rpl10a,ribosomal protein L10A,mouse, laboratory
Site of expression: EGFP is expressed primarily in layer 5b corticospinal, corticopontine pyramidal neurons, and small pyramidal cells.
Molecular note: Transgenic mice were generated using modified BAC Rp23-160M1. EGFP fused to the ribosomal protein L10a was inserted in the BAC in such a way that expression was driven by Colgalt2.