转红色荧光蛋白基因唐鱼外源基因拷贝数的测定  

Estimating the Copy Number of Exogenous Gene in Transgenic Tanichthys albonubes Expressing the Red Fluorescent Protein

作　　者：樊佳佳[1] 白俊杰[1] 马冬梅[1] 简清[1] 

机构地区：[1]中国水产科学研究院珠江水产研究所热带亚热带鱼类选育与养殖重点开放实验室,广东广州510380

出　　处：《生物技术通讯》2011年第1期61-65,共5页Letters in Biotechnology

基　　金：国家高技术研究发展计划(2009AA10Z105)

摘　　要：目的:测定转红色荧光蛋白基因唐鱼的外源基因拷贝数。方法:以杂合F5代和杂合F6代转红色荧光蛋白基因唐鱼为材料,以其外源插入片段(pDsRed-mylz2)与基因组插入位点5′侧翼区之间的边界序列作为特异性内参片段,同时以外源整合的红色荧光表达载体序列(pDsRed-mylz2)作为目的基因,采用实时荧光定量PCR技术测定外源基因整合的拷贝数。结果:运用外源基因与特异内参在同批次实时荧光定量PCR中的初始拷贝数比值,得到杂合F5代和杂合F6代转红色荧光蛋白基因唐鱼中插入的外源红色荧光表达载体序列pDsRed-mylz2的拷贝数均值均为3。结论:转红色荧光蛋白基因唐鱼的外源基因拷贝数为3。Objective：To estimate the copy number of exogenous gene in transgenic Tanichthys albonubes expressing the red fluorescent protein.Methods：The F5 and F6 generation heterozygous transgenic T.albonubes expressing the red fluorescent protein were used to estimate the copy number of exogenous gene by quantitative real-time PCR.The event-specific sequence was the border-sequence of pDsRed-mylz2 and 5’ flanking region sequence,the objective gene was pDsRed-mylz2.Results：The F5 and F6 generation heterozygous transgenic T.albonubes exogenous gene copy number were three by comparing the initial copy numbers between the event-specific sequence and the objective gene at the same quantitative real-time PCR.Conclusion：The exogenous gene copy number of transgenic T.albonubes was three

关 键 词：唐鱼 实时荧光定量PCR 外源基因 拷贝数 

分 类 号：Q78[生物学—分子生物学]