Cloning Of Elongation Factor-1A (EF-1A) Gene CDNA And Preliminary Function Analysis In Musa Acuminata

A method was used to screen the final target clone from banana fruit cDNA library by using 96-well plate and a pair of particular primer. A cDNA complete sequence of elongation factor la(EFl A) of banana with length of 1644bp had been obtained, and it was first time.The results of BLASTn and BLASTx for the obtained fragment showed that the sequence shared 84% homologous with Lilium longiflorum Thumb, 83% homologous with tobacco as well, it is conjectured that CDZ02 might be the gene encoding banana elongation factor 1 A(EF1A). Analysis of the ORF of the CDZ02 by the software of GENSCAN indicated that it contained an integrated ORF.RT-PCR was conducted to detect the expression of CDZ02 in different plant tissues of banana and the expression during different ripening stages of fruit. The procedure as follows: with the extracted RNA as model, the cDNA were reverse-transcribed, according to which a particular primer was designed afterwards, PCR was carried out with actin gene as a control.The gel image of RT-PCR was analyzed by the software - Scion image, it indicated that the elongation factor-1 alpha gene was expressed differently neither in different tissues of banana plant, nor during different ripening stages of post-harvest fruit. The expression of the gene were getting higher and higher in the fruit at the early stage of post-harvesting, which uptrend was either similar to ethylene release of post-harvested fruit, or to the intenerate course of fruit. The expression of the gene reached the highest spot on the 20th day after harvest, however it dropped to 50% on after 25 day. Furthermore, the gene was expressed in root with highest quantity, normal quantity in flower, the lowest in stem.