猪脂肪组织中UCP-2 mRNA表达产物的半定量分析  

Semiquantitative analysis of UCP-2 mRNA in fat tissue of pig by RT-PCR

作　　者：李民[1] 汪以真[1] 

机构地区：[1]浙江大学饲料科学研究所,浙江杭州310029

出　　处：《中国兽药杂志》2006年第1期20-23,共4页Chinese Journal of Veterinary Drug

基　　金：国家教育部新世纪人才基金(NCET-04-0543)

摘　　要：解偶联蛋白基因是新近发现的能够增加能量消耗,与脂肪代谢和能量调控密切相关的一组基因。本实验室建立了以18s rRNA为内标的RT-PCR法,半定量分析了猪脂肪组织中UCP-2 mRNA的表达水平。提取猪脂肪组织总RNA,经反转录和PCR扩增,PCR产物的电泳条带于VDS摄像系统下扫描灰度找出PCR线性扩增范围,确定RT-PCR的最佳循环数和Mg2+浓度。通过UCP-2 mRNA与18s rRNA PCR产物的灰度之比,即可计算出UCP-2 mRNA的相对含量。UCP genes are newly discovered genes that can increase the energy expenditure and involve the metabolism of fat and the regulation of energy. In this paper RT-PCR method was developed, which could be used for the semiquantitative analysis of UCP-2 mRNA in pig fat. 18s rRNA was used as interal standard. Total RNAs were extracted from fat tissue of pig. The mRNA was reverse-transcripted and the cDNA fragments of UCP-2 mRNA and 18s rRNA gene were amplified by RT-PCR using specific primers. The integrated optical density （IOD） was showed by scanning the electrophoresis bands of PCR products with VDS system. By studying the linear range of PCR amplification, the optimal cycle numbers and Mg^2＋ concentration were determined. The relative concentration of UCP-2 mRNA can be calculated by the IOD ratio of PCR products derived from UCP-2 mRNA and 18s rRNA.

关 键 词：猪 UCP-2 脂肪 RT—PCR 

分 类 号：Q786[生物学—分子生物学]