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Construction of Yeast Two-hybrid cDNA Library in Response to MNSV Infection in Melon |
WU Hui-jie,KANG Bao-shan,PENG Bin,LIU Li-feng,GU Qin-sheng* |
Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou 450009,Henan,China |
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Abstract In order to analyze the interaction between melon necrotic spot virus(MNSV)and host melon and screen the host factors interacted with the protein encoded by MNSV,this paper constructed cDNA library of melon infected with MNSV.During the cotyledon stage,melon variety ‘Baimei’which is susceptible to MNSV was inoculated with MNSV.After the plant showed typical necrotic spot symptom,the melon seedlings with remarkable symptom were collected.The double strand cDNA was synthesized by switching mechanism at 5′end of RNA transcript(SMART) technique.Then the cDNA was treated by duplex-specific nuclease(DSN).For the first time,the paper constructed the melon yeast cDNA library and evaluated the quality of this library.The library capacity was 4.5×106 cfu.Its titer was 3.6×107 cfu · mL-1.The library recombination rate was 100%.The size of inserting fragment of the library was 500-2 000 bp,indicating that the library had very good quality.Construction of this library has laid a foundation for analyzing interaction between MNSV and host,and understanding melon response to MNSV pathogenic mechanism.
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Received: 07 November 2019
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