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| Studies on Flanking Sequence of Potato Late Blight Resistance Gene by
Adapter Ligation-mediated PCR Amplifying |
| ZHANG Kun，XU Jian-fei，DUAN Shao-guang，PANG Wan-fu，BIAN Chun-song，LIU Jie，JIN
Li-ping* |
（Institute of Vegetables and Flowers， Chinese Academy of Agricultural Sciences， Key Laboratory of Biology and
Genetic Improvement of Tuber and Root Crop， Ministry of Agriculture， Beijing 100081， China） |
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Abstract Based on a potato（Solanum tuberosum L.）late blight resistance gene fragment
obtained by NBS profiling，specific nested primers were designed to amplify flanking sequences of the
fragment using adapter ligation-mediated PCR. The results indicated that，after digestion of genomic
DNA by restriction enzyme Dra Ⅰ，2 301 bp and 820 bp length sequence was amplified at the right and
left side of the known gene 536 bp fragment，respectively．Finally，the fragment was elongated 2 273
bp at right and 789 bp at left，which assemble a 3 443 bp full length sequence. A 2 413 bp candidate
gene including intron and extron was predicted with the software GENSCAN．A primer，RDZ，specific
to UTR region of the gene，was developed to amply resistant and susceptible genotypes．The results
showed a 2 571 bp fragment amplified by RDZ is linked to late b light resistance.
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Received: 21 May 2013
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