Construction of Full-length cDNA Library from Sugarcane Leaves and the Corresponding EST Analysis
In order to analyze gene expression pattern in the sugarcane(Saccharum Complex) leaves and set the basis for the cloning of some important functional genes, the functional leaves were taken as plant material from 3 development stage of sugarcane, and a full-length cDNA library was constructed by the method of Oligo-capping.After identification, the storage capacity of the library was 3.0×106 cfu, the recombinant rate was 89%, the full-length rate was 85%, and the average size of inserted fragment was about 1.0 kb.In total, two hundred and twenty-eight efficient ESTs(expressed sequence tags) of 5' end were obtained from this cDNA library.Bioinformatics analysis showed that these ESTs were assembled into 154 TUTs(tentative unique transcript).The homologous analysis of the 154 TUTs on NCBI showed that these 87 TUTs(including 117 ESTs) were determined to be the tags of gene with putative function, these genes were found to be involved in the biological process including cell development, signal transport, protein synthesis, transcription, stress tolerance response, energy metabolism and so on.