通过PCR技术简单扩增奶牛高GC含量的rDNA重复序列  

Simple Amplification of GC-Rich Repetitive Sequences in Bos taurus rDNA with PCR

作　　者：王波[1] 唐冬生[1] 蒋泓[1] 洪亚辉[2] 萧浪涛[3] 周天鸿[1] 

机构地区：[1]暨南大学生命科学技术学院,广东广州510632 [2]湖南农业大学生物科学与技术学院,湖南长沙410128 [3]湖南农业大学湖南省植物激素与生长发育重点实验室,湖南长沙410128

出　　处：《湖南农业大学学报（自然科学版）》2005年第6期602-604,共3页Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金资助项目(30170737);广东省自然科学基金资助项目(04011645)

摘　　要：为了探索利用PCR简单扩增高GC含量的长重复序列的方法,经反复摸索后选用LAPCR法即LAPCRTaq酶结合GC缓冲液Ⅱ来扩增奶牛高GC含量的长片段重复序列,成功获得了全长2538bp,GC含量为65.7%的DNA序列,其中ITS1的GC含量高达80%.探索了克隆复杂DNA序列时引物设计的特别规则、反应体系的改进、DNA聚合酶的选用等措施.In order to develop a simple method to PCR amplification of the long high GC content repetive sequence, the LA PCR method with LA PCR Taq DNA polymerase and GC buffer II was used to amplify the rDNA sequences in Bos taurus after repeat experiment. The sequences of rDNA in Bos taurus were gained, whose size is 2 538 bp, and GC content as high as 65.7% including GC content of its ITS1 as high as 80%. The conditions for amplifying high GC content repetitive sequences such as the special rules of primer design, the improvement of reaction system, the selection of DNA polymerase were discussed.

关 键 词：高GC含量 重复序列 核糖体DNA 聚合酶链反应 奶牛 

分 类 号：Q7[生物学—分子生物学] S823[农业科学—畜牧学]