Construction Of Suppressive Subtractive Library Of Dairy Cattle X-, Y-Sperm

It has been a long concern to control the gender of newborn livestock and the sex control of the dairy cattle is especially demanding in today. The separation of semen is considered to be a much easier and lower cost approach in various ways of sex control. In this paper, to find the meaningful differences between X and Y sperms, differences of this two types of sperms in the transcriptome has been studied according to the separation mechanism of HY antigen-antibody.Transfer RNA, as the intermediate carrier of genetic information, plays an important role in the process of spermatogenesis, sperm survival, sperm-egg bonding. In this study, the extraction of RNA in cattle sperm has been explored according to the structure of the sperm and characteristic of the RNA content. The RNA from mammalian sperm has been extracted creatively using the of extraction methods for plant tissues and the integrity of the extracted RNA was proved by testing the production of PCR amplification of gene GAPDH. The extracted X-sperm, Y-sperm transcripts have also been amplified at the same proportion for the lower sperm RNA content using SMART technology and the amplified cDNA was digested by the restriction endonuclease Rsaâ… . The, digested cDNA was divided into two parts, which connected to different connectors, and hybrided with the unconnected drivers for two cycles. Then after two rounds of PCR amplification, the production was connected to the pGEMTeasy carrier and transformed into the body of E. colicoliform. The sequences data from 366 clones have been obtained in the library and 292 in the anti- library were compared through BLAST after removing the carriers. It was found that most of the sequences were not found the homologous genes. This may be due to the imperfect of the cattle EST. Thus the database could be enriched by the research of the new ESTs and theoretical basis for experiment of differences in protein might also be found.