Background/Objectives: The Berlin Fat Mouse Inbred line 860 is a model for juvenile obesity. Previously, a recessive major effect locus (jObes1) on chromosome 3 between 34 and 44 Mb has been found to be responsible for 39% of the variance of total fat mass at 10 weeks in a (BFMI860 x C57BL/6NCrl) F 2 population. The aim of this study was fine mapping of the jObes1 locus.Subjects/Methods: An advanced intercross line (AIL) was generated from the initial F 2 mapping population. Three hundred and forty-four male mice of generation 28 were excessively phenotyped and genotyped using the MegaMuga mouse chip containing 22 164 informative single-nucleotide polymorphisms. Expression of candidate genes was investigated in gonadal adipose tissue, liver and whole brain from mice of different genotype classes. Classical genetic complementation tests were performed to test candidate genes. Results: The high mapping resolution of the AIL reduced the confidence interval for jObes1 from 10 to 0.37 Mb between 36.48 and 36.85 Mb. This region was highly significantly (logarithm (base 10) of odds (LOD) score after Benjamini and Hochberg correction (LOD (BH))>50) associated with total fat mass starting at puberty (6 weeks). Male homozygous carriers of the jObese1 BFMI allele had 3 g more fat than the other genotypes. Surprisingly, this genotype class showed lower body mass until weaning at 3 weeks (LOD (BH) =3.2). The mapped interval contains four genes. Bbs7, the most likely candidate gene that also caused obesity in the complementation test was differentially expressed in all tissues examined, whereas the neighboring cyclin A2 (Ccna2) gene showed differential expression in gonadal adipose tissue. Conclusions: Using an AIL, the confidence interval for jObes1 could be 27-fold reduced by finding chromosomal recombinations. Although Bbs7 is the most likely obesity gene in the jObes1 region, neighboring genes cannot be entirely excluded. Further examinations are needed to enlighten the mechanism leading to physiological consequences on body mass and fat mass in juvenile animals.