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Bulletin of Botanical Research ›› 2013, Vol. 33 ›› Issue (6): 713-717.doi: 10.7525/j.issn.1673-5102.2013.06.013

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Construction and Evaluation of Yeast Two Hybrid cDNA Library of Banana Leaf Infected by BBTV

TAN Lao-Xi;ZHANG Yu-Liang;ZHOU Peng;ZHANG Shao-Yan;WANG Jian-Hua;ZHANG Xiu-Chun;LIU Zhi-Xin*   

  1. 1. Key Laboratory of Biology and Genetic Resources of Tropical Crops,Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Science,Haikou 571101;2.College of Environment and Plant Protection,Hainan University,Haikou 570228
  • Received:1900-01-01 Revised:1900-01-01 Online:2013-11-20 Published:2013-11-20
  • Contact: LIU Zhi-Xin
  • Supported by:
     

Abstract: BBTV is a major limiting factor for banana production. In this report, we presented a BBTV infected banana (family Musaceae) leaf cDNA library constructed by using a kit named Make Your Own MatePlateTMLibrary System. Total RNA was extracted from BBTV-infected banana leaves by using modified CTAB method. The first stand cDNA was synthesized by reverse transcription of mRNA with SMART technique. To generate cDNA library, the cDNA was digested with SfiI and then sub-cloned into SfiI side of pGADT7 vector. The recombinant plasmids were transferred into E.coli via electroporation. The result of the plasmids from amplified cDNA library showed that the library contained 2.0×106 independent clones and the size of most inserts were 700-2 000bp in the library. The recombination rate of library was 87.5%. These results indicated that the library was suitable for screening proteins interacting with BBTV coded protein. It will facilitate the investigation of interaction between BBTV and the host.

Key words: BBTV, banana leaf, Yeast-two-hybrid cDNA library

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