Anorexia anx/anx mutant mice

Anorexic anx/anx mice Jackson Lab strain 000624.

Mutation mapped to a 0.2 cM interval residing between the markers D2Mit133 and Jojo5 chromosome 2 (Chr 2: bp 118, 889, 896–120, 175, 1081) (Lindfors et al., 2011).

Possible genes affected

Lindsfor 2011: NADH dehydrogenase (ubiquinone) 1a-subcomplex (Ndufaf1)

Kim 2017: tyrosine kinase receptor Tyro3 which they conclude is not the anx-mutation but a strain specific modifier of anx-phenotypes

Tyro3 Genotyping

TYRO3 protein tyrosine kinase 3 (Tyro3) mutation in anx/anx eliminatees a NlaIV restriction site.

• primers 5′-GATGGCGCTGAGGCGGAGCATG and 5′-CGCGGC- CGGAGGTCTGGCAG ; annealing temperature 72C

• subsequent digestion with NlaIV

• wildtype -> 175 bp fragment, anx/anx -> 250 bp product

Genotyping 1998 mice

• 2024-1-17 Tyro3 primers ordered from FSU BIO Core

• 2024-1-18 NlaIV ordered from NEBiolabs, along with 50bp ladder

PCR: 95C for 1min, 72C for 2min, 30 cycles