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Construction and Characterization of a cDNA Library from Wheat Infected with Fusarium graminearum Fg 2
Department of Plant Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada
General Commission of Agricultural Scientific Research (GCASR), Department of Biotechnology, P.O. Box 113 Douma-Damascus, Syria
* Author to whom correspondence should be addressed.
Received: 26 November 2010; in revised form: 14 January 2011 / Accepted: 17 January 2011 / Published: 18 January 2011
Abstract: Total RNA from wheat spikes infected with F. graminearum Fg2 was extracted and the mRNA was purified. Switching Mechanism at 5' end of the RNA Transcript (SMART) technique and CDS Ill/3' primer were used for first-strand cDNA synthesis using reverse transcriptase by RT-PCR. Primer extension polymerase chain reaction was used to construct the double-strand cDNA that was digested by proteinase K, then by Sfi I and fractionated. cDNAs longer than 0.5 kb were collected and ligated to λTriplEx2 vector followed λ phage packaging reaction and library amplification. The qualities of both unamplified and amplified cDNA libraries were strictly checked by conventional titer determination. One hundred and sixty five plaques were randomly picked and tested using PCR with universal primers derived from the sequence flanking the vector. A high quality cDNA library from wheat spikes that have been infected by F. graminearum was successfully constructed.
Keywords: 15 acetyl deoxynivalenol (15ADON); 3 acetyl deoxynivalenol (3ADON); cDNA library; Fusarium graminearum Fg2; Triticum aestivum
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Al-Taweel, K.; Fernando, W.G.D.; Brûlé-Babel, A.L. Construction and Characterization of a cDNA Library from Wheat Infected with Fusarium graminearum Fg 2. Int. J. Mol. Sci. 2011, 12, 613-626.
Al-Taweel K, Fernando WGD, Brûlé-Babel AL. Construction and Characterization of a cDNA Library from Wheat Infected with Fusarium graminearum Fg 2. International Journal of Molecular Sciences. 2011; 12(1):613-626.
Al-Taweel, Khaled; Fernando, W. G. Dilantha; Brûlé-Babel, Anita L. 2011. "Construction and Characterization of a cDNA Library from Wheat Infected with Fusarium graminearum Fg 2." Int. J. Mol. Sci. 12, no. 1: 613-626.