Difference Analysis Of DNA Methylation Level On Rongchang Pig

Rongchang barrow at different tissues(liver,heart and psoas) and different weight stages(10kg, 20kg,35kg,50kg,80kg,100kg) of DNA methylation level were detected by HPLC(High Performance Liquid Chromatography).The purpose was to investigate DNA methylation level change rule Rongchang barrow of different tissues and different weight stages,also to state mechanism of epigenetic regulation in the process of growth and development and offer further basic data on.The results were as follows:1The establishment and optimization of HPLC technical systemRNase T1 using purified DNA,was the hydrolysis of a single deoxy-nucleoside mixture after high-temperature solution chain in P1 nuclease and bovine intestinal alkaline phosphatase,contrast to the deoxy-nucleoside of 2'-deoxycytidine(dC),2'-deoxyadenosine(dA),2'-deoxyguanosine(dG), 2'-deoxythymidine(dT),5-deoxycytidine(5-~mc) standard mixture and determine parallel test,the parallel groups were of no difference.It came to optimize the chromatographic conditions:the symmetryshieldTM RP18 column,a mobile phase of phosphate acid 0.1%(v/v),waters,2487 UV detector at 273nm,the column temperature was 25℃,the sensitivity was 2.0000.DNA requirements for only 20μL.The samples were separated completely in 20min,The method was proved simply and reliably.2.The change tendency of genome DNA methylation level at different tissues and weight stagesThe results showed that:the DNA methylation level showed a descending trend with weight increase.DNA methylation level in the stages of 10kg were significantly higher than that of 20kg, 35kg,50kg,80kg and 100kg(p＜0.01).In the stages of 20kg,it was significantly higher than that of 80kg,100kg(p＜0.01).In the stages of 35kg,50kg,80kg,the levels were significantly higher than that of 100kg(p＜0.01).There were no differences among other weight stages(p＞0.05).It is concluded that this DNA methylation level bears good structure selectivity and relativity, the psoas was y=-0.000241x+0.0346(R~2=0.4677),the liver was y=-0.000174x+0.0419(R~2=0.5679), the heart was y=-0.000155x+0.0363(R~2=0.3514).Different tissues,weight,the interaction between different tissues and weight of DNA methylation level were significant(p＜0.01).The impact of order was different tissues(η~2=0.395)＞ weight(η~2=0.393)＞the interaction(η~2=0.157).The descending of different tissues of DNA methylation level order was psoas＞heart＞liver.