The Basic Study On The Fat-1Transgenic Cattle

ω-3PUFAs have been shown to play an important role in health. Enriched with ω-3polyunsaturated fatty acids modulate expression of a number of genes with such broad functions as cell proliferation, growth and apoptosis and cell signaling and transduction, these effects, seem to regulate coronary artery disease, hypertension, atherosclerosis, psychiatric disorders and various cancer. However, human and mammals can not synthesize ω-3PUFAs by themselves, so they obtain ω-3PUFAs by dietary intake. Fat-1gene encode ω-3PUFAs desaturase, it play importance role in the synthesis of ω-3PUFAs, which can convert ω-6PUFAs into ω-3PUFAs, then increase the content of ω-3PUFAs in the body. The study was transferred the fat-1gene into bovine genome and then production the transgenic cattle by animal clothing technology. It is importance to detect transgene copy number and influence to the genome gene, which benefit to know the transgenic cattle. Realtime PCR and cDNA microarrays were employed to detect the transgene copy number and influence to the genome gene. The results were as follows:1. Detection of positive transgenic miceThe PCR was used to initial identify two transgenic cattle firstly, then the southern blot was employed to further validated the PCR result. The result showed that the two methods obtained consistent result and the two detected cattle were transgenic positive, can processed follow-up experiment.2. Detection of transgene copy numberThe study was used the absolute quantitative real-time PCR to detect the copy number. The parameter of the standard curve was:Log2N=-3.16Ct+40.76(R2=0.9942). The copy number of two cattle are4.62±1.03,5.46±1.26respectively.3. The influence of the expression of genomic geneThe study was used the gene expression profiling, transgenic cattle as detect group and wild type as control group to fluorescent labeling and hybridize with chip in the meantime, through analyze the ratio of fluorescent intensity of the two groups with probe to detect the change of gene expression. The result showed that Fat-1transgenic cattle and wild-type cattle have43653discrepancy expressed transcripts according to the Agilent software. This study we choose differentially expressed genes of p-value≤0.05and fc≥1to analysis. The result was demonstrated that to change the composition of polyunsaturated fatty acids can effects on gene expression, and then affect the physiological activity and pathological process through different mechanisms.