A study on polymorphism of exon and intron 2 of leptin gene and detection of SNPs in Mahabadi goat using PCR-SSCP

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Abstract

Leptin, a 16-kDa protein secreted from white adipocytes, has been implicated in the regulation of feed intake, energy expenditure, and whole-body energy balance, reproductive, immunity, growth  and obesity gene product (obese) coded by a gene called ob gene. The present study has been designed to determine polymorphism in exon and intron 2 region of leptin gene in Mahabadi goat using PCR-SSCP. Blood samples were collected from150 Mahabadi goat belong to the Department of Animal Science, University of Tehran (Karaj). DNA was extracted using salting out method. DNA quality and quantitative was determined using spectrophotometer and 1% agarose gel. A fragment 271bp of exon and intron 2 region of leptin gene was amplified using the polymerase chain reaction. SSCP technique was used to analyze polymorphism in this segment of DNA. PCR products were run in 12% polyacrylamide gel electrophoresis for 21h. Gels were stained with silver nitrate. Developing pattern showed the same pattern in all animals and was monomorph. The results of this research showed seven mutations for this locus that the Mahabadi goats breed for these mutations are monomorph and these mutations ordinary was reported in other breeds of goats.

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