半定量RT-PCR检测猪延髓中色氨酸羟化酶2基因表达方法的建立  

Detection of mRNA Gene Expression of Tryptophan Hydroxylase-2 in Pig Medulla Oblongata by Semi-quantitative RT-PCR

作　　者：马红娜[1] 邓衔柏[1] 马勇江[1] 何敏[2] 习欠云[3] 宁章勇[1] 张桂红[1] 卢培成[1] 张曼玉[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]广东科贸职业学院,广东广州510430 [3]华南农业大学动物科学学院,广东广州510642

出　　处：《动物医学进展》2010年第7期1-6,共6页Progress In Veterinary Medicine

基　　金：国家973计划项目(2009CB118805)

摘　　要：利用半定量逆转录-聚合酶链反应(RT-PCR)技术,测定仔猪延髓组织中色氨酸羟化酶2(TPH-2)基因mRNA表达水平,以β-actin为内标,分析相对表达量及其与驱赶应激的关系。通过探讨和优化PCR体系中退火温度、Mg2+浓度和循环次数等参数,建立了检测延髓组织中色氨酸羟化酶-2基因mRNA表达的半定量RT-PCR体系。通过分析色氨酸羟化酶-2 PCR产物与β-actin PCR产物电泳后的灰度比值,可知驱赶应激对TPH-2基因mRNA表达量的影响。The gene expression of tryptophan hydroxylase-2（TPH-2）in pig medulla oblongata was detected by RT-PCR method.The relationships between relative gene expressions of TPH-2 and hustle stress were analyzed by using β-action as internal standard.The semi-quantitative RT-PCR system detecting the mRNA gene expressions of TPH-2 was set up by discussing and optimizing Mg2＋ concentration,anneal temperature and the number of loop.In experiment,the ratio of integrated optical density（IOD） of TPH-2 PCR and β-actin PCR were analyzed.The result indicated that the influence of hustle stress on gene expression of TPH-2 may be known.

关 键 词：色氨酸羟化酶2 逆转录-聚合酶链反应 应激 

分 类 号：Q789[生物学—分子生物学] Q554.7