Construction and Characterization of Yeast Two-Hybrid cDNA Library Derived from Roots of Gossypium barbadense Inoculated with Verticillium dahliae

doi:10.13560/j.cnki.biotech.bull.1985.2014.12.017

Abstract

Abstract: A yeast two-hybrid cDNA library derived from roots of Gossypium barbadense cv. Pima90-53 inoculated with severe virulence Verticillium dahliae was constructed based on SMART^IM technique and homologous recombination reaction. Detection of the library indicated that its capacity and titer were 2.82×10⁶ and 5.02×10⁸ cfu/mL, respectively. The result from colony PCR showed that the length of insert cDNA fragments ranged from 500 to 1 500 bp on average and the recombination rate was 93.33%. These results demonstrate that the library meets the demands of the standard cDNA library, which will be useful for screening interaction proteins, determining signal transduction components and analyzing the structure and function of disease-resistant proteins in the future.

Key words: Verticillium dahliae, Gossypium barbadense, Yeast two-hybrid, cDNA library

Yang Jun,Zhang Yan,Wang Weiqiao,Rong Wei,Wang Xingfen,Ma Zhiying. Construction and Characterization of Yeast Two-Hybrid cDNA Library Derived from Roots of Gossypium barbadense Inoculated with Verticillium dahliae[J]. Biotechnology Bulletin, 2014, 0(12): 105-110.

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