Microarray Profiling For Differential Gene Expression In Ovarian Follicles Of Pigs Between Erhualian And Large White

Litter size is one of the important factors which impact the benefit of porcine industry. In this research, we used gene chip to analyze the differences of genes expression about ovarian follicles between Large White(LW), which have strong reproduction, and erhualian pigs which have opposite characters. According to the above analysis and genes'physiological and biochemical function, we choose BAX as candidate, isolated and identificated it used comparative genomics and bioinformatics, studied their tissue and space-time expression differences used real-time PCR, and then detected SNPs within them used direct sequencing method. Finally, the association analysis had been done between the polymorphisms and the reproductive performance in female Large White, Chinese lean type new line DIV populations. The main results are as following:1. We used swine whole genome Affymetrix oligo chip contained 23937 probes to detect the genes differentially expressed in ovarian follicles between Large White and erhualian pigs. According to multiple comparison, we screened related genes that had statistical significance (P<0.05). Taking a fold change (FC) of two or greater and the significance level as the criteria, a total of 133 transcripts showed differential expression. A set of 29 transcripts such as BMPR1B, H3F3A, MT-2B,ELA2belonged to the up-regulated group, and another set of 104 transcripts such as GALP, KIT, BAX, P53 belonged to the down-regulated group in Chinese Erhulian sows.2. This experiment choose 8 genes, including P53, BMPR1B, CNN2, ITM2A, WIT1, H3F3A, LOC396850, GALP, utilization of fluorescence quantitative real time polymerase chain reaction (RT-PCR) to validate reliability of the results of gene chips. The results show the expression tendency of mRNA of 7(except CNN2)genes in 2 sorts of data of RT-PCR and gene chips are the same and their correlation are significance, thereby indicating that the results of gene chips are credible.3. (1) A 416bp DNA sequence of pig BAX gene was isolated; (2) For BAX gene:a PCR-Earâ… -RFLP assay was established to detect the C/T; (3) Association analysis for the C/T with reproductive traits was preformed in three independent populations. Statistical analysis demonstrated that Earâ… polymorphism was associated with NBA in the first of Large White sows, an additive effect was detected (P<0.05 or P<0.01).