Cloning, Selecting, Identification And Characterization Of The Differentially Expressed Genes Between Mongolian Sheep Ovary

The trait of prolificacy of sheep is an important approach to increase more economic benefit, in order to research the molecule genetic mechanism of fertility character of Mongolian sheep and other sheep breeds at home and aboard, we take Mongolian sheep as our main research object, adopting techniques such as molecule genetic markers as well as SSH. Starting with candidate genes and selecting different genes. We Studied moleculer genetic Polymorphism of Mongolian sheep and other sheep breeds, and discussed the factors and molecule genetic mechanism of multiparous biparous sheep:1 .Study of candidate genes on sheep prolificacyBy the methods of PCR-SSCP and RFLP, we studied polymorphism on 5 sites of BMPRIB, BMP 15 gene and GDF 9 genes from 7 sheep breeds, 375 individuals, such as Mongolian sheep, Hulunbeier, Chinese Merino, Suffolk, Dorset, German Merino and Inner Mongolian fine wool sheep .The results showed that (1) the genotype of BMP15 gene exon 1 was delected in four domestic breeds and German Merino, three genotypes were displayed the deletion of 3 base pairs in 28-30, causes the deletion of tenth amino acid, the other two breeds have not any variation. (2). the genotype of BMPRIB gene has the same variations as Booroola Merino (A746G), and the other five breeds have not any variation. (3). The analysis of PCR-SSCP shows that G8 sites of GDF9 Gene didn't existed any variations in seven sheep breeds .2,Identication and Characterization of the differentially expressed Genes between Mongolian sheep ovary(1) we constructed forward and reverse subtracted cDNA libraries between ovaries from multiparous single and biparous Mongolian sheep by suppression subtractive hybridization(SSH) technique successfully, thus, 384 clones were randomly selected from each cDNA subtracted library, and identified by PCR, obtaining 768 masculine clones,The Length of the sequences was between 150bp～1000bp.(2) Using forward subtractive cDNA and reversal subtractive cDNA as Probe,in the subtracted cDNA library, multiparous biparous is tester and multiparous single Mongolian sheep is driver, 47 clones were selected by blot dot technology , through sequencing and blast, they represent 10 ESTs of sheep, known in sheep, 4 of them are unknown but 27 of them have high similarity with human and other species genes, and 6 have no obvious similarity with known genes. 3,Cloning and sequence analysis Mongolian sheep full length BMP15 geneFull length BMP15 gene was cloned in Mongolian sheep, including 2 exons and 1 intron, full-length DNA 6642 bp, ORF 1182bp, coding 393 aa, by sequence analysis we found it has a high similarity with human and other species genes, but it has a less similarity with fish and birds, about 20%. At the same time, we also forecaste the advanced structure of BMP15...