Cloning, Polymorphism And Expression Patterns Analysis Of Porcine LPL Gene

Lipoprotein lipase (LPL) gene is a multifunctional protein, which catalyses the hydrolysis of the triglycerides component of Chylomicrons and very low density lipoproteins (VLDL), regulates the fatty metabolism, and facilitates the integration and absorption of lipoprotein by the cell. So it is considered as one of the key enzymes in fat deposition. This study was designed to clone the full-length coding sequence of Yorkshire by RT-PCR, to predict its structural and functional domains with informatics method, to analyze its polymorphisms in five pig breeds including wild boar, min pig, Yorkshire, Landrace and Duroc by PCR-SSCP, and to characterize its tissue expression pattern in Min pig at 180-day-old of age by real-time PCR. The results are as follows:1. The full-length coding sequence of LPL gene of Yorkshire, composed of 1436 bp encoding 478 amino acids, was cloned. The structural and functional domain was initially predicted by bioinformatics methods. And the homology of LPL gene from different species was compared and the phylogenetic tree was constructed with Neighbor-Joining method.2. Several SNPs and a insertion/deletion mutation were found in exon 5 and 10. Population genetic analysis showed that there were significantly different in the distribution of genotypes in 5 five breeds.3. The tissue expression pattern of LPL gene at min pig (180 days-old) was constructed by Real-time PCR. The results showed that LPL gene was unevenly expressed in all 13 tissues detected including heart, liver, stomach, spleen, kidney, lung, large intestine, small intestine, uterus, ovary, dorsal muscle, lower Limb muscle and fat with high level in heart, dorsal muscle, lower Limb muscle , fat and low level in uterus and ovary.