Lab Anim Res.  2016 Mar;32(1):70-73. 10.5625/lar.2016.32.1.70.

Rapid and efficient identification of the mouse leptin receptor mutation (C57BL/KsJ-db/db) by tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) analysis

Affiliations
  • 11Laboratory Animal Center/Department of Medical Genetics, College of Medicine, Hallym University, Chuncheon, Korea. jgsuh@hallym.ac.kr
  • 2Institute of Natural Medicine, Hallym University, Chuncheon, Korea.

Abstract

The C57BLKS/J-Lepr(db) mouse has a point mutation in the leptin receptor gene and is one of the most useful animal model for non-insulin dependent diabetes mellitus in human. Since the homozygote of C57BLKS/J-Lepr(db) mouse is infertile, detection of point mutation in the leptin receptor gene is important for efficient maintaining strains as well as mass production of homozygotes. To develop a rapid and efficient genotyping method for C57BLKS/J-Lepr(db) mouse, the tetra-primer amplification-refractory mutation system polymerase chain reaction (ARMS-PCR) was used. The 407 and 199 bp PCR products were amplified from normal (+/+) mice; while the 407 and 268 bp PCR products were amplified from homozygotes (db/db) mice; and the 407, 268, and 199 bp PCR products were amplified from heterozygotes (db/+) mice. This result showed that the tetra-primer ARMS-PCR analysis by us is suitable to detect point mutation of the leptin receptor gene. Taken together, our method will dramatically reduce animal use for maintenance of strains as well as production of homozygote in the C57BLKS/J-Lepr(db) strains.

Keyword

Leptin receptor; genotyping; db mouse

MeSH Terms

Animals
Diabetes Mellitus
Heterozygote
Homozygote
Humans
Leptin*
Mice*
Models, Animal
Point Mutation
Polymerase Chain Reaction
Receptors, Leptin*
Leptin
Receptors, Leptin
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