The targeting vector has been generated using C57BL/6J DNA and transfected into TaconicArtemis C57BL/6NTac ES cell line.
Exon 1 contains the translation initiation codon.
Exon 5 has been flanked by loxP sites (size of loxP-flanked region: 0.9 kb).
The conditional KO allele has been generated after Flp-mediated recombination by crossing chimeras to a Flp-Deleter on a C57BL/6 background.
The conditional KO line has been derived using C57BL/6NTac animals and the Flp-transgene was removed by segregation.
The constitutive KO allele can be generated by Cre-mediated recombination. Deletion of exon 5 should result in loss of function of the Mgea5 gene by deleting both active sites of the catalytic domain (nucleophile: amino acid 175 and proton donor: amino acid 177) and by generating a frameshift from exon 4 to exons 6-9 (premature Stop codon in exon 6).
The remaining recombination sites are located in non-conserved regions of the genome.
According to NCBI database, a gene is called E330018D03Rik (Entrez ID: 320319) is located within Mgea5 gene (intron 10 to intron 11). E330018D03Rik is located approx. 10 kb downstream of Mgea5 exon 5 in the same orientation.
Datamining and Design performed in 2010.
Gene Family: Transferase
Genetic Background: C57BL/6
Gene Symbol: Mgea5
Gene Accession Number: NM_023799.3
Allele Type: cKO
Availability: Cryopreserved at Taconic
Official Gene Name: meningioma expressed antigen 5 (hyaluronidase)