Expression and structure of an elongation factor-1 alpha gene (MeEF1) from cassava (Manihot esculenta Crantz)

• Published in: Euphytica 2001-01, Vol.120 (1), p.49-58
• Main Authors: Suhandono, S, Hughes, J, Brown, K, Hughes, M
• Format: Article
• Language: English
• Subjects: elongation factor-1^a; Hordeum vulgare; Manihot esculenta; MeEF1 gene
• ISSN: 0014-2336
• DOI: 10.1023/A:1017543318423

In order to isolate an elongation factor-1 alpha (EF-1 alpha ) gene from cassava, a lambda EMBL3 genomic library, made from a single cassava genotype (MBRA 534, from the CIAT cassava germplasm collection), was screened with a full length EF-1 alpha cDNA clone (blt63) from barley. Six positive clones were isolated from an amplified library and 4866 bp from one clone (MeEF1) were subcloned into pGEM3zf(-) and pGEM5zf(-). The sequence has 2709 bp 5' of the translation start site, 1347 bp of coding sequence split into two exons by an intron (374 bp) and 435 bp 3' of the stop codon. A 657 bp intron was also found 5' of the translation start site. The coding sequence is very similar, with 86% DNA sequence identity and 95% deduced amino acid sequence identity, to an EF-1 alpha gene from Arabidopsis thaliana. The promoter region of MeEF1 contains 3 putative control elements that are located upstream of the transcription start site. These control elements include a TEF1 box, a TELO box and two TATA boxes. The gene is expressed in early stages of development and in young tissue. Transient expression using particle bombardment shows that the promoter drives uidA gene expression in leaves of cassava and Arabidopsis. An interesting feature of the MeEF1 gene is that the presence of the 5'UTR intron affects the level of expression.