TAIL-PCR的技术改良及对香蕉NBS-RGC5基因侧翼序列的克隆  

Modification of TAIL-PCR and NBS-RGC5 gene flanking sequence cloning from banana

作　　者：罗静瑶[1,2] 陈云云[1,2] 谢俊[1,2] 韦双双[2] 夏幽泉[2] 汤华[1,2,3] 

机构地区：[1]海南大学海南热带生物资源可持续利用国家重点实验室培育基地,海南海口570228 [2]海南大学农学院,海南海口570228 [3]热带作物种质资源保护与开发利用教育部重点实验室,海南海口570228

出　　处：《广东农业科学》2013年第10期143-145,170,共4页Guangdong Agricultural Sciences

基　　金：国家自然科学基金(30860149);教育部科学技术研究重点项目(210172);教育部热带作物新品种选育工程中心与作物学重点学科联合项目(lhxm-2012-2)

摘　　要：香蕉枯萎病是一种毁灭性真菌病害,对香蕉的种植和产业发展危害严重,研究香蕉枯萎病相关的抗性基因,具有重要的理论意义。对TAIL-PCR进行技术改良,以简并引物组合的方式与特异性引物进行扩增,代替最初的单一简并引物方式,在TAIL-PCR第一轮扩增的大循环中设置不同的退火温度;采用改良后的TAIL-PCR成功克隆了巴西蕉NBS型抗性蛋白(NBS-RGC5)基因的侧翼序列。Fusarium wilt disease is a destructive fungus disease for banana, it has serious negative impact on banana planting and industrial development, so it is significant to research the resistant gene related to banana wilt disease. In this article, part of gene sequence of Nucleotide Binding Site type of Resistant Gene （NBS-RGC5） from Brazil Banana （Musa acuminate ssp. Cavendish AAA）was isolated using modified TAIL-PCR method. Compared with the original protocol, there were two main modifications of the TAIL-PCR .. （1） Instead of the single of the degenerate primer, the combination of two degenerate primers was used in the modified TAIL-PCR.（2）The annealing temperature in the 15 super cycles in the Primary PCR was set differently. The final results demonstrated that the modified of TAIL-PCR was an instant and efficient method to clone the flanking sequences from known region.

关 键 词：香蕉 枯萎病 热不对称交错PCR(TAIL-PCR) NBS-RGC5 侧翼序列 

分 类 号：Q758[生物学—分子生物学]