莱航鸡rDNA重复基因家族的克隆  

Cloning on Complicated Repetitive Sequences of rDNA Genes Family in Leghorn Chicken

作　　者：唐冬生[1] 李芳[1] 张勇[1] 蒋泓[1] 胡大林[1] 张细权[2] 李月琴[3] 周天鸿[3] 

机构地区：[1]佛山大学,广东佛山528000 [2]华南农业大学动物科学学院,广东广州510642 [3]暨南大学生命科学与技术学院,广东广州510632

出　　处：《中国家禽》2007年第3期13-16,共4页China Poultry

基　　金：国家自然科学基金(30671194);广东省自然科学基金(04011645);广东省科技攻关项目(2006B20101012)资助项目

摘　　要：动物rDNA基因是一种GC含量较高、结构复杂的重复序列。通过结合生物信息学技术,经反复摸索后选用LAPCR法扩增莱航鸡rDNA基因重复序列,经测序鉴定最终克隆了莱航鸡的3个rDNA基因及其2个间隔序列。研究对克隆复杂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、循环参数的调整等进行了探索。The rDNA genes in animals are the kind of repetitive sequences with high GC content and complicated structure. Combining the bioinformatics technique,the repetitive DNA sequences of rDNA family of Leghorn chicken were gained. The LA PCR method with LA PCR Taq DNA polymerase and GC buffer was used to amplify the complicated repetitive sequences of rDNA genes in Leghorn chicken after repeat experiment. Furthermore,the PCR products were cloned into pMD18-Simple T cloning plasmid. After sequencing identification three genes and two internal transcribed spacers （ITS1 and ITS2） of rDNA had been cloned. The measures for cloning the complicated DNA sequences such as the special rules of primer design,the improvement of reaction system,the selection of DNA polymerase and the adjustment of cycle parameter had been explored in this study.

关 键 词：RDNA基因 基因克隆 莱航鸡 

分 类 号：Q959.233[生物学—动物学]