Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (9): 1696-1702.doi: 10.3864/j.issn.0578-1752.2012.09.004

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Construction and Sequences Analysis of Full-Length cDNA Library of Tobacco Leaves

 GONG  Da-Ping, JIE  Min-Min, SUN  Yu-He   

  1. 1.中国农业科学院烟草研究所/中国农业科学院烟草遗传改良与生物技术重点开放实验室，山东青岛266101
  • Received:2011-10-20 Online:2012-05-01 Published:2011-12-21

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Abstract: 【Objective】 The full-length cDNA library from tobacco leaves mRNA was constructed. EST data were generated and these gene families of development, metabolizability and resistance were detected and ananlyzed. 【Method】 The Cap-trapper method was optimized to constructe a full-length enriched cDNA library of tobacco seedling leaves. The assembly and annotation of EST data were completed by bioinformatics program. 【Result】 A high quality full-length cDNA library was constructed successfully from tobacco leaves. The titer was 1.2×106 pfu•mL-1 and the average length of inserted cDNA fragments was 1.4 kb. A total of 5 280 clones were sequenced from the library, and 5 233 expressed sequence tag (EST) sequences were obtained. The ESTs were assembled into 3 922 unigenes including 564 contigs and 3 358 singletons. The comparative analysis shows that approximately 89.7% of the unigenes have homologs in other plants. These genes were involved in cell development, signal transduction, transcription and synthesis, stress tolerance response, energy metabolism and so on. Nicotine synthesis and transform, and disease resistance genes were identified. 【Conclusion】 A high-quality full-length cDNA library of tobacco leaves was successfully constructed by Cap-trapper method. Meanwhile, development, nicotine synthesis and transform, stress resistance and disease resistance were identified by sequences analysis of EST.

Key words: tobacco, full-length cDNA library, EST, function annotation

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