Construction of a cDNA library of the leaf of Leymus chinensis and analysis of partial expressed sequence tags

Abstract

Abstract: Total RNA of Leymus chinensis (“Jishengyihao”) leaves was isolated by TRIZOL reagent and a cDNA library was constructed using SMART technology. The primary library had a high titer of 10⁶ cfu/mL, in which 98% of the clones were recombinant and the insert cDNAs were from 0.5 kb to 3.0 kb. The amplified library had a titer of 10¹¹ cfu/mL. The positive signals of TC and γ-TMT gene were detected by PCR of the amplified library. High quality sequences were shown by 307 of the cDNA clones.In the cDNA library, 117 clones were non-redundant, and BLAST led to the identification of several putative genes (e.g. glyceraldehyde-3-phosphate, RNase S-like protein precursor, translation elongation factor 1, translation initiation factor 1A, chloroplast psbA gene for D1 protein). This high quality cDNA library provides a useful tool for further study of the molecular mechanisms of the secondary metabolism of vitamin E and of gene expression in L. chinensis.

CLC Number:

WANG Li-juan, JIN Zhi-ping, WANG Neng-fei, LI Xiao-feng, LIU Gong-she. Construction of a cDNA library of the leaf of Leymus chinensis and analysis of partial expressed sequence tags[J]. Acta Prataculturae Sinica, 2009, 18(1): 65-71.

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