Clonng And SNP Detection Of Plin Gene And Its Assocaition Analysis With Crass And Fat Traits In Duks

Perilipin(PLIN) plays an important role in the development of fatness traits. This study is to find out the relationship among hereditary variation of PLIN carcass and fat traits. The current research adopted Peking duck as material.The coding sequence of PLIN and 3'-RACE sequence were successfully obtained by RT-PCR and analyzed by bioinformative method.The hereditary variation between Peking duck(Z2,Z4,Z2×Z4) at the 8th week after birth and Cherry Valley duck by PCR-RFLP and DNA sequencing was detected. The regularity of distribution was compared respectively in different group. The relativity of hereditary variation of PLIN and carcass and fat traits was analyzed in order to discover notable genetic markers for major economic characters. The main results achieved in this study as follows:1. Cloning and bioinformative analyses of PLINThe coding sequence(CDS) and 3'-UTR of PLIN gene were successfully obtained, which was 2212bp.GenBank was assigned which accession number was NC₀07319. The ORF contains 1590 nucleotide, including initiator codon(ATG) and terminator codon(TGA). The cDNA sequence have 8 extrons. A amino acid sequence with 530aa was deduced by the cDNA sequence. Homologous comparison was done between the corresponding section in human(NM₀01145311) and chicken(NM₀01127439), and the homology were 87% and 81%. The structural and functional forecast of PLIN protein shows that the protein molecular weight is 57.23KDa, and its isoelectric point is 9.23. Furthermore, the lyophobic maximum and minimum of PLIN protein are located at 273th and 429th aa respectively. PLIN protein have two Low components complexity which were at the region of 188-200aa and 213-235aa. The conserved domain of PLIN protein was built by 5-365 amino acid residues, whose structure character accord with the characteristic of PAT family.2.Genetic diversity of different extrons and 2th intronThe hereditary variation was detected between Peking duck(Z2,Z4,Z2×Z4) and Cherry Valley duck by PCR-RFLP and DNA sequencing. The result showed, there is no polymorphic in 3th,4th,5th,6th,7th, and 8th extrons. Moreover, a same sense mutation was found in extron 2(Câ†'T); a missense mutation was found in intron 2(Câ†'T).3. Correlation analysis among polymorphism and carcass and fat traits The Least-square linear correlation analysis among polymorphism of extron 2 carcass and fat traits showed as follows. In Cherry Valley duck, there was extremely significant difference between CC and CT (P<0.01) in carcass weight, carcass net weight, the percentage of abdomen fat weight (AFW); There was significant difference between CC and CT(P<0.05) in AFW and the percentage of SWF; and there was no significant correlation between three genotypes in the rest carcass and fat traits. In Z4, carcass net weight was significantly different between TT, CC and CT (P<0.05); and there was no significant correlation between three genotypes in the rest carcass and fat traits. In Z2×Z4, carcass net weight was significantly different between CC and CT (P<0.05), and there was no significant correlation between three genotypes in the rest carcass and fat traits. In Z2, TT was not detected;and there was no significant correlation between three genotypes in all the carcass and fat traits(P>0.05).The Least-square linear correlation analysis among polymorphism of intron 2, carcass and fat traits showed as follows:In Z4, there was extremely significant difference between EE and DE(P<0.01) and significant difference between EE and DD(P<0.05) in carcass net weight; there was no significant correlation between three genotypes in the rest carcass and fat traits. In Z2XZ4, there was extremely significant difference between DD and DE(P<0.01) in breast muscle weight and significant difference between DD and EE(P<0.05) in SFW; there was no significant correlation between three genotypes in the rest carcass and fat traits. In Z2 and Cherry Valley duck, there was no significant correlation between three genotypes in all the carcass and fat traits (P>0.05).PLIN gene of Beijing duck was initially cloned successfully in this study, including the complete coding region and 3'untranslated region, which provided a theoretical basis for the regulatory function of PLIN gene in the lipid metabolism and the duck meat. SNP loci of PLIN gene of four duck species were detected and the correlation among the PLIN gene mutation locis, carcass and fat traits were analyzed. Significant correlation was founded, which has a certain theoretical and practical reference value to carry out the cultivation of superior varieties.