磁珠富集法分离菠萝蜜(Artocarpus heterophyllus)微卫星标记及系列分析  

Isolation of microsatellite in jackfruit (Artocarpus heterophyllus) by magnetic beads

作　　者：王艳红[1] 李映志[1] 丰锋[1] 叶春海[1] 

机构地区：[1]广东海洋大学农学院,广东湛江524088

出　　处：《果树学报》2010年第6期1046-1051,共6页Journal of Fruit Science

基　　金：国家自然科学基金(30771481);广东省自然科学基金(07010103);广东省科技攻关计划(2007020200006)

摘　　要：利用磁珠富集法分离出菠萝蜜微卫星位点,并成功设计出菠萝蜜(Artocarpus heterophyllus Lam.)SSR(简单序列重复,Simple sequence repeat)分子标记引物。基因组DNA经MseI酶切后,用生物素标记的简单重复序列做探针与其杂交,杂交复合物固定到包被有链霉亲和素的磁珠上,经过一系列的洗涤过程,含有SSR的酶切片段被吸附到磁珠表面。这些片段经洗脱下来后,先用对应的引物扩增,再进行克隆和测序。实验共挑出83个阳性克隆进行测序,成功设计合成SSR引物19对。并通过这19对SSR引物对20份菠萝蜜种质资源进行分析,全部能扩增出特异性条带,其中18对引物扩增的条带具有多态性,可以作为菠萝蜜分子标记使用。Genomic DNA of Jaekfruit （Artocarpus heterophyllus Lam.） was extracted and digested with restriction enzyme Mse 1. The fragment of DNA were hybridized with biotins labeled Oligo （GA）10 , （A C）10 probes and several other probes, and the hybridized eomplex was fixed by magnetic beads coated with streptavidin. The selected fragments were amplified using adapter specific primers and cloned into pGEMT-vcctors and transtbrmed into Ecoli hosts. An SSR enriched genomie DNA library was thus constructed. 83 clones were selected by colony -PCR method and then sequenced. And 19 pairs of SSR primers were found usable.

关 键 词：菠萝蜜 微卫星 磁珠富集法 

分 类 号：S667.8[农业科学—果树学]