实验用西藏小型猪CYP3A46基因扩增梯度PCR方法的建立  

Establishment of gradient PCR amplification method for CYP3A46 gene of laboratory Tibet minipigs

作　　者：卢康荣[1] 白静 张嘉宁 谢水林[2] 顾为望 黄黎珍[2] 王万山 LU Kang-rong1, BAI Jing2 ,ZHANG Jia-ning3, XIE Shui-lin2, GU Wei-wang3, HUANGLi-zhen2, WANG Wan -shan3(1.Basic Medical College of Southern Medical University, Guangzhou Guangdong 510515, China;2.School of Bioscience and Bioengineering, South China University of Technology, Guangzhou Guangdong 510006, China;3. Laboratory Animal Service Center, Southern Medical University, Guangzhou Guangdong 510515, Chin)

机构地区：[1]南方医科大学基础医学院,广东广州510515 [2]华南理工大学生物科学与工程学院,广东广州510515 [3]南方医科大学实验动物中心,广东广州510515

出　　处：《科技视界》2018年第19期88-89,共2页Science & Technology Vision

基　　金：广东省科技计划项目(2011B060300028;2012B040304010)

摘　　要：目的:建立西藏小型猪CYP3A基因梯度PCR扩增方法。方法:以西藏小型猪肝脏总cDNA为模板,基于CYP3A46基因引物,对CYP3A46进行梯度PCR扩增(设计温度梯度55-65℃)。PCR产物进行琼脂糖凝胶电泳,获得特异性好的最佳退火温度。结果:各退火温度均能特异的扩增出1500bp左右的目的条带。结论:本反应体系及各温度梯度均可特异性扩增西藏小型猪CYP3A46基因,为进一步克隆表达奠定了基础。Objective ： Establishing gradient PCR amplification method for CYP3A46 gene of Tibet minipigs. Methods ：Based on CYP3A46 primers, using total c DNA of Tibet minipig liver as template, CYP3A46 was amplified by gradient PCR, temperature gradients were set as 55-65 ℃. PCR fragments was checked by DNA agarose gel electrophoresis,trying to get the best annealing temperature. Results ： A specific band of 1500 bp corresponding to the target band could be amplified by any temperature from 55 to 65 ℃. Conclusions ： CYP3A46 gene of Tibet minipigs could be specifically amplified by this reaction system and temperature gradients, laying a foundation for the further cloning and expression.

关 键 词：细胞色素P450酶 西藏小型猪 克隆 

分 类 号：Q78[生物学—分子生物学]