The Study On Cloning, Localization, Polymorphism And Tissue Distribution Of Porcine IGFBP6 And CYR61 Gene Fragments

IGFBP6 (insulin-like growth factor binding protein 6) and CYR61 (cysteine rich angiogenic inducer 61) are members of the IGFBP superfamily which can combine with IGFs. They regulate growth development and metabolism by regulating the binding capability between IGFs and IGFs receptor. In this study, IGFBP6 and CYR61 were selected as candidate genes for growth and fat metabolism in the pig. The fragments cloning, chromosomal localization, tissue distribution, prokaryotic expression, polymorphism identification and association with production traits, provided useful information on its application in marker assisted selection (MAS) in the future. The main results are as follows:1. Full-length sequence of coding region of the porcine IGFBP6 gene and partial sequence of the CYR61 were cloned using the pig specific primers which were designed from the pig EST contig obtained by the method of comparative genomics according to the corresponding human cDNA sequence.2. The somatic cell hybrid panel (SCHP) and the INRA-University of Minnesota porcine radiation hybrid (IMpRH) panel were used for determining the chromosomal location of the pig IGFBP6 and CYR61 genes. The PCR results were analyzed using the mapping analysis tools available on the web server. The IGFBP6 gene was mapped to the region of SSC5p11-p15, with close linkage to SW439 (LOD = 5.43); The CYR61 gene was mapped to the region of SSC4q21-q24, with close linkage to SW1461 (LOD = 5.43).3. Tissue distribution was analyzed by the sequencing and the RT-PCR in 9 different tissues including heart, liver, spleen, lung, kidney, muscle, adipose tissue, stomach and small intestine from female Landrance and 6 different tissues including heart, liver, spleen, lung, kidney and muscle from male Landrance × Large White hyrid pig. The IGFBP6 gene was detected in all tissues. CYR61 gene was detected in heart.4. IGFBP6 Taa I -RFLP (T-C) polymorphism in exon 4 was demonstrated by sequnencing and PCR-RFLP. The polymorphism for this mutation was analyzed in ten different pig breeds. This SNP is polymorohic in all breeds.5. The Taa I -RFLP (T-C) of IGFBP6 gene was analyzed in the experimental pig population including Tongcheng, Landrace, Large White , Landrace ♂×(Large White × Tongcheng) ♀ and Large White ♂× (Landrace × Tongcheng) ♀. The association of the...