Molecular Cloning And Analyzing Of Obese Gene CDNA In Pecking Duck

Obese gene (ob) is a gene cloned recently. It encodes a protein hormone. The product of ob gene-Leptin is an important signal reflecting body fat mass and controlling weight. It plays a key role in the control of food intake and energy expenditure. Leptin can remarkably cut down the amounts of fat mass, stimulate development of the adolescence % regulated the beginning of procreate and the function of blood made. To exploit and take advantage of the leptin, the Pecking duck obese gene was cloned and analysis by RT-PCR and the alignment of ob gene in Pecking duck and some other animals was contrasted.The total RNA was abstracted from the Pecking duck adipose tissue and the liver tissue. Consulting the common sequence of mouse and human, the primer was devised on the sequencing data of chicken mRNA.The Pecking duck ob gene was amplified by RT-PCR technology. The duck ob gene is 463bp.Retrieve the fragment and link it with the vector-PUC18. Transform the fragments into the Ecoli DH5 cell. To scalping the positive clone by blue and white spot and abstract the plasmid DNA from the cell. To electrophone plasmidDNA in 1% agrose e gel. The result indicated that the goal fragments have cloned to the vector. The length of the insert fragments is similarly to the goal fragments. These indicated that the successful of the clone of Pecking duck protein encoded sequence.The result of determine sequence shows that Pecking duck ob gene is 463bp and it encodes 135 amino acids and have a frame reading. Contrast the alignment of ob cDNA sequence between Beijing duck and other animals. The alignment of of ob cDNA sequence between Pecking duck and huma, rat, pig, chicken, cattle, goat is 81 percent, 91 percent, 81 percent, 95 percent, 83 percentand 83 percent respecetively. The alignment of Leptin protein sequence between Pecking duck and human, rat, pig, chicken cattle, goat is 67 percent, 77 percent, 62 percent, 80 percent, 65 percent and 64 percent respectively.