Related Analysis Of Renin And IL-10 With Porcine Gestation Length

The gestation length serves as one of the most importent reproductive traits. Reducing the gestation length,taking swine breeding into account,directly results in a shorter reproductive cycle;correspondingly reduce the production cost by increasing the reproductive efficiency of sows.Gestation length is a quantitative trait which is affected by several genes' cooperation.Due to the amount of genes and every single effect it provides,as well as environmental factors,regular breeding methods have scarcely any apparent genetic improvement on this character.The QTL related with gestation length is located at the 1,9,15 chromosome in sus scrofa.According to swine- human genome comparison and both physiological and biochemical function,two candidate genes renin and interleukin-10 have been selected in this experiment.In this experiment several methods like bioinformatics and conservation of linkage with swine and man are being applied to clone these genes and polymorphic sites have been made in order to analyze the relativity with the gestation length and and total number of piglet born.1.Based on swine's EST which has a high homology with human renin gene and the cDNA of other species like mice,five primers have been designed on the sequences with great similarity within several animals.Meishan's cDNA has been chosen as the template which is sequenced and cloned.After splicing and integrating,the cDNA, 1333 base pair,has been acquired including the whole CDS and parts of 5' and 3' untranslationed region.Later,the DNA sequence has amplified,cloned and sequenced which was examined as 13332 base pairs.2.With the help of software Clustal X,ORF Finder,Spidey,MEGA4.0,PSORTâ…¡, renin's gene structure,the structure of protein,property,the phylogenetics tree,has been predicted and analyzed.We find that this gene with 9 extrons and 8 introns matching the GT-AG rule,is extremely resemble with rumen animals,less similar with rodents,and least conformity with primate.3.Relied on cDNA of IL-10 gene,seven primers have designed and the whole gene has been cloned with 4062bp.After analyzing the structure of the gene,4 introns have been included and complied with the GT-AG rule.4.RT-PCR has been used to exam renin expression in different tissues,the results demonstrate that kidney,ovaries,oviduct have expressed this gene with the highest one founded in kidney. 5.Different DNA of Qingping,Large white,Landrace and Duroc have been amplified, cloned and sequenced.Transvertion of C/T at the 6 intron,288bp,and A/G at the 7 intron 114bp in renin as well as the transvertion A/G in the third intron of IL-10 at 835bp have been found based on sequences comparison.Appropriate PCR-RFLP SNP typing methods have been established.6.The mutation in the sixth intron of renin can be cut by enzyme Hhaâ… .Qingping,New Qingping,large white,landerace,Duroc and Chinese lean meat pig strain Dâ…£₁,Dâ…£₂,altogether seven kinds of swine have been typed.The genetype frequency and gene frequency were analyzed in order to dig out the relativity with gestation length and and total number of piglet born.The result shows that the specific site with genetype TT plays more prominent effect in foreign pigs,whereas local breed and cultivated strain has significant variation from foreign swine even achieved the extremely significant variation.The dominant effect on gestation length in this site in Qingping has reached significant level as well.In the Dâ…£₁ strain,hogs' gestation length with gene type CC were significantly longer than that of TT and CT gene type. The dominant effect and the additive effect on total number of piglet born in this site in all the groups are not significant.7.The mutation in seventh intron of renin can be cut by enzyme bstxâ… .Qingping,New Qingping,large white,landerace,Duroc and Chinese lean meat pig strain Dâ…£₁,Dâ…£₂,altogether seven kinds of swine have been typed.The genetype frequency and gene frequency were analyzed in order to dig out the relativity with gestation length and total number of piglet born.The result shows that allele gene A has a more conspicuous advantage in any swinery,besides duroc.The gene frequency has significant variation and the dominant effect on gestation length in Qingping was significant,0.93day.The additive effect on total number of piglet born in this site in Duroc has reached significant level,hogs' total number of piglet born with gene type GG were significantly more than that of AG and AA gene type.8.The mutation in third intron of IL-10 can be cut by enzyme Eco81â… .Qingping,New Qingping,large white,landerace,Duroc and Chinese lean meat pig strain Dâ…£₁,Dâ…£₂,altogether seven kinds of swine have been typed.The genetype frequency and gene frequency were analyzed in order to dig out the relativity with gestation length and total number of piglet born.The result shows that the specific site with gene type GG plays more prominent effect in Qingping,New Qingping and duroc,whereas within large white,landerace,Dâ…£₁,and Dâ…£₂ AA acted a more important role.Most swinery has significant variation from each other.The dominant effect on gestation length in this site in duroc has reached significant level,consequently pigs with gene type AG have a longer gestation length than those of AA gene type.The additive effect on total number of piglet born in this site in large white has reached significant level,hogs' total number of piglet bom with gene type GG were significantly fewer than that of AG and AA gene type.