Porcine Obesity Gene 5 'regulatory Region Sequence Determination And Preliminary Analysis

Leptin ,an adipocyte-derived signal ,is product of the obesity gene(OBS)and is important in the regulation of body weight,body fat mass,and food intake.Elucidation of the mechanisms underlying modulating this gene has significance for the swine breeding and increasing efficiency of lean meat tissue.For the first time ,16kb large fragment,namely,intro I ,exon I ,5'region of ob gene,was restrictive analyzed and sequenced,and found two novel microsatellites in intro I .The study mainly included Primary analysis of BAG plasmid clone which contains porcine obese gene fragment. In the study, Primers designed according to a known 6000bp fragment of porcine obese gene were used to amplify the BAG clone. It was confirmed that the BAG clone contains the Ob gene by sequencing the PCR product.Restriction analysis of the BAG clone was carried out by Pulse-Field-Gel-Electrophoresis. PCR products which had been labeled using DIG were used to locate the region of obese gene in BAG clone. DNA fragments generated by restriction enzyme, which were derived from BAG clone, were subcloned into PGEM-7zf. Pieces of 5' region of obese gene were detected and sequenced , and a 16.8Kb large fragment was analyzed by both restriction analysis and sequencing.