半定量RT-PCR法分析猪肝脏中铜锌超氧化物歧化酶mRNA表达水平  

Assay of the Copper Zinc Superoxide Dismutase mRNA Level in Pig Liver by Semiquantitative RT-PCR

作　　者：胡迎利 徐春兰[2] 汪以真[2] 

机构地区：[1]河南省郑州牧工专科学校,河南郑州450008 [2]浙江大学饲料科学研究所教育部分子营养重点实验室,浙江杭州310029

出　　处：《中国兽药杂志》2006年第4期20-24,共5页Chinese Journal of Veterinary Drug

基　　金：河南省重大攻关项目-无公害猪用饲料添加剂的研究与开发(03222011100)

摘　　要：细胞内铜锌超氧化物歧化酶(CuZnSOD)是动物体内一种重要的抗氧化酶。本实验室构建了优化的半定量RT-PCR法,以18S rRNA为内标,研究猪肝脏组织中CuZnSOD基因mRNA表达水平。提取猪肝脏组织总RNA,经反转录后进行扩增,先寻求PCR线性扩增范围,确定RT-PCR的最佳循环数和Mg2+浓度。扩增后用VDS摄像系统扫描PCR扩增产物的电泳条带灰度。通过CuZnSOD PCR产物的灰度与18S rRNA PCR产物的灰度之比,即可计算出CuZnSOD mRNA的相对含量。Copper zinc superoxide dismutase （CuZnSOD） is an important antioxidase in animals. This experiment set up a RT-PCR process that could be used for semiquantitative analysis CuZnSOD mRNA. 18S rRNA was used as interal standard. Total RNA was extracted from liver tissue of pig. The mRNA was reverse-transcripted and the cDNA fragments of CuZnSOD and 18S rRNA gene were amplified by RT-PCR using the gene specific primers. The integrated optical density （IOD） was showed by scanning the eletrophoresis bands of PCR products with VDS system. By studying the range of PCR products linearity amplification, the optimal cycle number and Mg2^2＋ concentration were made certain. The relative concentration of CuZnSOD mRNA could be calculated by the IOD ratio of CuZnSOD and 18 S rRNA

关 键 词：铜锌超氧化物歧化酶(CuZnSOD) RT-PCR 肝脏 猪 

分 类 号：Q503[生物学—生物化学]