腺病毒介导的MAT2B过表达促进猪肌内脂肪细胞分化 赵存真;李黄琨;马云;杨公社 【期刊名称】《畜牧兽医学报》 【年(卷),期】2017(048)005 【摘 要】The aim of this study was to explore the effect of MAT2B on porcine intramuscular preadipocyte differentiation by constructing overexpression vector in vitro mediated by adenovirus.A pair of exclusive primers was designed according to the GenBank sequence information of MAT2B gene,and MAT2B was amplified by PCR.The obtained PCR products were inserted into a shuttle vector pAdTrack-CMV,then the gene was identified by sequencing.The results showed that shuttle vector (pAdTrack-CMV) and backbone vector (pAdEasy-1) were in homologous recombination,and the adenovirus recombinant vector (pAd-MAT2B) was constructed successfully.Then the recombinant adenovirus DNA was digested by the Pac I,the pAd-MAT2B adenovirus was produced,and we used it to infect the porcine preadipocytes.The expression level of mRNA and protein of MAT2B significantly increased.Oil Red O staining assay showed that overexpression of MAT2B promoted lipid accumulation in intramuscular preadipocyte.Moreover,the expression of adipogenic gene PPARγ and aP2 mRNA were significantly promoted when infected by Ad-MAT2B.The results suggest that MAT2B is overexpressed in expression vector in vitro mediated by adenovirus,and MAT2B play a positive role during porcine intramuscular preadipocyte differentiation.%本研究旨在通过构建腺病毒介导的体外超表达载体,探究腺苷甲硫氨酸转移酶(MAT2B)对猪肌内脂肪细胞分化的影响.本研究以猪脂肪组织为试验材料,提取总RNA,并反转录得到cDNA,参考GenBank收录的猪MA T2B基因mRNA序列设计引物,PCR扩增并连接到腺病毒穿梭载体pAdTrack-CMV,进行测序鉴定.结果表明,构建的穿梭载体与骨架载体pAdEasy-1可以实现同源重组,即腺病毒重组载体pAd-MAT2B构建成功;用Pac Ⅰ限制性内切酶酶切线性化pAd-MAT2B载体并回收质粒大片段转染293A细胞可以实现病毒的成功包装,病毒滴度测定可满足原代细胞侵染需要.转染猪原代肌内脂肪细胞后,MA T2B的mRNA和蛋白水平实现了显著的上调.油红O染色结果表明,过表达MA T2B促进了肌内脂肪细胞脂质积累;实时定量结果证明,MAT2B促进成脂标志基因PPARγ和aP2表达的显著上调.综上所述,腺病毒介导的体外表达载体可以成功的实现MAT2B基因超表达;MAT2B正向调控猪肌内脂肪细胞分化. 【总页数】8页(P802-809) 【作 者】赵存真;李黄琨;马云;杨公社 【作者单位】信阳农林学院牧医工程学院,信阳464000;信阳农林学院牧医工程学院,信阳464000;信阳师范学院生命科学学院,信阳464000;西北农林科技大学动物脂肪沉积与肌肉发育实验室,杨凌712100 【正文语种】中 文 【中图分类】S828;S813.1 【相关文献】 1.莱芜猪和杜洛克猪肌肉H-FABP基因表达量与肌内脂肪和脂肪酸含量关联分析 [J], 呼红梅;王继英;郭建凤;张印;沈彦锋;武英 2.猪肌内脂肪前体细胞与皮下脂肪前体细胞分化过程中基因差异表达分析 [J], 张罕星;朱晓彤;束刚;周桂炫;高萍;高淑静;张常明;江青艳;陈瑶生 3.过表达MAT2A基因促进猪肌内脂肪细胞分化的研究 [J], 赵存真;吴海港;唐雪峰;焦凤超;马云;刘纪成 4.慢病毒介导shRNA干扰MAT2A与MAT2B基因抑制猪肌内脂肪细胞分化 [J], 赵存真; 易本驰; 陈培荣; 李建柱; 赵云焕; 朱忠珂 5.二花脸猪与大约克猪生长期肌内脂肪合成与水解基因表达特征的比较研究 [J], 高勤学;李俊;刘红林;王林云;徐银学 因版权原因，仅展示原文概要，查看原文内容请购买 本文来源：https://www.wddqw.com/doc/9b014e364bd7c1c708a1284ac850ad02de8007e7.html