Cloning of LEC1 gene from embryogenic callus and its expression analysis during somatic embryogenesis in longan

LEC1 gene was cloned from embryogenic callus and its expression patterns were determined during somatic embryogenesis in longan(Dimocarpus longan Lour.).The RT-PCR(reverse transcription polymerase chain reaction),RACE(rapid amplification of cDNA ends) and TAIL-PCR were used to isolate the complete cDNA sequence of LEC1 from longan embryogenic callus,and the bioinformatics methods were used to analyze the cDNA sequence and putative amino acid sequence.The full length LEC1 cDNA,which had been submitted to the DDBJ/EMBL/GenBank database(the accession number was GU584089.2),was 803 bp,in addition,the putative protein had 222 amino acids,and the identity to the other polypeptides varied between 35%-70%.Finally,the qRT-PCR(real-time quantitative transcription PCR) method was used to determine the mRNA transcription levels of the gene.LEC1 was expressed at the different stages during somatic embryogenesis,showing a single peak curve with the relative higher mRNA transcription level of LEC1 gene at the heart embryo stage and the torpedo embryo stage.