PurH基因在大蒲莲猪及其杂交后代肌肉中表达及其与肌苷酸含量关联分析

Correlation between PurH Gene Expression and Inosine 5’-monophosphate(IMP) Content in Dapulian and its Hybrid Combination Pigs

旨在研究PurH基因在大蒲莲及其杂交组合猪肌肉中的表达差异,并与肌苷酸含量进行相关性分析,为探讨肌苷酸代谢基因PurH与重要风味物质的关系奠定理论基础。选择95.0 kg体重大蒲莲猪(DPL)、长白×大蒲莲猪(LDPL)、杜洛克×长白×大蒲莲猪(DLDPL)各8头,屠宰后取背最长肌,提取总RNA。应用荧光定量PCR(qPCR)对11个常用的候选内参基因和PurH基因表达量进行检测,同时利用高效液相色谱测定肌肉中肌苷酸含量。结果表明:(1)geNorm软件评价分析显示,YWHAZ和RPL4是肌肉中表达最稳定的内参基因,可利用其表达量的几何均值标准化PurH基因表达量;(2)肌肉中肌苷酸含量依次为DPL>LDPL>DLDPL,DPL分别比DLDPL和LDPL提高25.35%(P<0.05)和15.93%(P>0.05),DPL、LDPL肌肉中PurH基因表达量分别比DLDPL增加50.00%和25.00%(P<0.05),DPL肌肉中PurH基因表达量比LDPL增加20%(P>0.05);(3)DPL、LDPL、DLDPL背最长肌中肌苷酸含量与PurH基因表达量变化趋势一致。相关性分析发现,肌苷酸含量与PurH基因表达量显著正相关。研究揭示,肌肉中PurH基因mRNA表达量和肌苷酸含量依次为DPL>LDPL>DLDPL,而且PurH基因表达量与肌肉肌苷酸含量呈显著正相关,为培育富含肌苷酸的优质猪肉提供理论支撑。

The study was aimed to analyze relative expression level of PurH and the correlation between gene expression and inosine 5'-monophosphate(IMP)content of longissimus dorsi in Dapulian pigs and its hybrid combination pigs.This research revealed the correlation between PurH gene and flavor substances.Eight Dapuian(DPL)pigs,eight Landrace×Dapulian(LDPL)pigs and eight Duroc×Landrace×Dapulian(DLDPL)pigs weighting 95.0 kg were chosen.Total RNA was isolated from the longissimus dorsi after slaughter using Trizol reagent.The expression of the eleven candidate genes and PurH gene were evaluated using real time quantitative PCR(qPCR),IMP content in longissimus dorsi was detected by HPLC.The results showed that:(1)The geNorm program showed that YWHAZ and RPL4 were the most stable reference genes in muscle,and the geometric mean of YWHAZ and RPL4 expression could be used to standardize PurH gene expression;(2)IMP content of longissimus dorsi in Dapulian(DPL)pigs was higher than Landrace×Dapulian pigs(LDPL)and Duroc×Landrace×Dapulian pigs(DLDPL),IMP content of longissimus dorsi in Landrace×Dapulian pigs was higher than Duroc×Landrace×Dapulian pigs.Compared with DLDPL,IMP content in DPL significantly increased by 25.35%(P<0.05).Although IMP content in DPL was 15.93%higher than that in LDPL,but the difference was not significant(P>0.05).The expression level of PurH gene in DPL and LDPL respectively remarkably increased by 50.00%and 25.00%than that in DLDPL(P<0.05),there was no significant difference in the expression of PurH gene between DPL and LDPL,although the expression of PurH gene in DPL increased by 20%than LDPL;(3)The change of IMP content of longissimus dorsi in DPL,LDPL,DLDPL was consistent with the expression of PurH.ADSL gene relative expression and IMP content were significantly correlated.The mRNA expression of PurH and IMP content in longissimus dorsi was the highest in DPL,with LDPL the second highest and DLDPL the lowest,and they were significantly correlated,which provided a theoretical support for producing IMP rich high quality pork.