RAMP与REMAP分子标记技术及其应用  

RAMP and REMAP Molecular Marker System and Their Application

作　　者：王明霞[1] 柳李旺[1,2] 龚义勤[1] 赵丽萍[1] 李培[1] 黄丹琼[1] 

机构地区：[1]南京农业大学作物遗传与种质创新国家重点实验室,南京农业大学园艺学院 [2]北达科他州立大学植物科学系,法戈,美国,58105

出　　处：《分子植物育种》2006年第6期859-865,共7页Molecular Plant Breeding

基　　金：国家自然科学基金项目（30300238,30571193）;江苏省自然科学基金项目（创新人才BK2004418）;江苏省高技术研究计划资助项目（BG2005314）;上海市科技兴农重点攻项目（沪农科攻字2004第9-1号）资助.

摘　　要：引物设计是RAMP和REMAP标记分析的关键,二者都使用锚定的微卫星序列引物。目前已开发出多个RAMP锚定微卫星序列引物,与不同的RAPD引物进行扩增,得到更多的引物组合。REMAP使用反转录转座子的外向LTR引物和锚定微卫星序列引物。而RAMP分析采用不对称PCR循环,在PCR扩增的后15个循环中,退火温度设为2个;REMAP则采用常规PCR循环。扩增产物可在聚丙烯酰胺凝胶、变性聚丙烯酰胺凝胶、琼脂糖凝胶上进行电泳分离,检测时可使用同位素、银染或溴化乙锭染色等技术。目前,这两种标记技术已在作物和动物种质资源的遗传多样性分析、遗传连锁图谱构建、转基因植株基因稳定性分析等方面都有成功的应用。The primer design is a key step for RAMP and REMAP, in which anchored simple sequence repeat primers were used. Many anchored microsatellite primers were developed for RAMP, and more primer combinations could be obtained by combining with different RAPD primers. Primers facing outward from LTRs in retrotransposon and anchored microsatellite primers were designed for REMAP. An asymmetric PCR profile was employed with setting of one high and one low annealing temperature for RAMP-PCR during the last 15 cycles, and REMAP-PCR program was as usual. The amplicons can be separated by polyacrylamide gel, denaturing polyacrylamide gel or agarose gel, and detected by autoradiography, silver or EB staining. RAMP and REMAP had been successfully applied in germplasm identification and genetic diversity analysis, genetic map construction and genomic stability analysis of transgenic plants in many crops and animals.

关 键 词：RAMP REMAP 分子标记 

分 类 号：Q943.2[生物学—植物学]