花生TRAP-PCR分析体系的建立  

Establishment of TRAP-PCR Analytic System in Peanut

作　　者：徐磊[1] 王晶珊[1,2] 隋炯明 王晓杰[1] 乔利仙[1] 

机构地区：[1]青岛农业大学生命科学学院,山东青岛266109 [2]青岛市主要农作物种质创新与应用重点实验室

出　　处：《青岛农业大学学报（自然科学版）》2010年第1期42-45,51,共5页Journal of Qingdao Agricultural University(Natural Science)

基　　金：国家自然科学基金(30871544);公益性行业科研专项(nvhyzx07-014);青岛农业大学博士启动基金(630731)资助

摘　　要：目标区域扩增多态性(target region amplified polymorphism,TRAP)是通过一个依据EST序列设计的固定引物和一个针对外显子或内含子特点设计的随机引物对开放阅读框(open reading frames,ORFs)进行扩增。本研究首次建立了适于花生TRAP分析的PCR扩增体系:在15μl总反应体系中,模板DNA50ng,引物浓度1.0μmol/L,dNTPs浓度0.25mmol/L,Taq DNA聚合酶1U。利用该体系对24个花生品种DNA进行扩增,得到了清晰稳定的条带,且这些条带具有一定的多态性,说明所建立的体系可用于花生遗传多样性分析。Target region amplified polymorphism （TRAP） aims for the amplification of open reading frames （ORFs） by using a fixed primer which is designed from the targeted EST sequence in the database and an arbitrary primer which is an arbitrary sequence with either an AT - or GC - rich core to anneal with the intron or extron, respectively. The TRAP- PCR system was set up firstly in peanut. The optimal system was in 15 ul reaction volume containing about 50ng template DNA, 1.0umol/L of each primer, 0.25mmoL/L of dNTPs, IU Taq polymerase. DNAs of 24 peanut varieties were amplified using the system. The result showed that the amplified products were clearly and stable, and some of them were polymorphic, indicating that the analytic system was suitable for the diversity detection in peanut.

关 键 词：花生 目标区域扩增多态性(TRAP) 分析体系 

分 类 号：S565.2[农业科学—作物学] Q943.2[生物学—植物学]