Molecular cloning and characterization of a novel microsomal oleate desaturase gene DiFAD2 from Davidia involucrata Baill
In the conversion of oleic acid to linoleic acid, δ12-fatty acid desaturase (δ12-FAD) is involved. Based on the conserved oligo amino acid residues of the FAD2 genes from other plants, a new full-length cDNA (DiFAD2) encoding a δ12-FAD was cloned from Davidia involucrata Baill. Sequence analysis indicated that the DiFAD2 gene had an open reading frame (ORF) of 1 149 bp, coding for 382 amino acids residues of 44.3 kDa, pI of the deduced protein was 8.8. The deduced amino acid sequence of the cloned DiFAD2 showed high identities to those genes of other plant δ12-FAD. RT-PCR showed that DiFAD2 was expressed in all tissues and expression was abundant in young stems. Expression of DiFAD2 is not enhanced by low temperature and the altered polyunsaturated fatty acid content in leaves treated with low temperature may be due to the post-transcriptional regulation of the DiFAD2 gene or the other FAD2 gene family regulation.
Additional key wordsamino acid residues open reading frame Saccharomyces cerevisiae
Davidia involucrata δ12-fatty acid desaturase
- Di18S rRNA
Davidia involucrata 18S rRNA
open reading frame
polymerase chain reaction
rapid amplification of cDNA ends
reverse transcriptase-polymerase chain reaction
polyunsaturated fatty acids
Unable to display preview. Download preview PDF.
This work was supported by the national infrastructure of national resources for science and technology (2005DKA21403).
- McCartney, A.W., Dyer, J.M., Dhanoa, P.K., Kim, P.K., Andrews, D.W., McNew, J.A., Mullen, R.T.: Membranebound fatty acid desaturases are inserted co-translationally into the ER and contain different ER retrieval motifs at their carboxy termini. — Plant J. 37: 156–173, 2004.CrossRefPubMedGoogle Scholar
- Niu, B., Ye, H.X., Xu, Y., Wang, S.H., Peng, S.M., Ou, Y.C., Tang, L., Chen, F.: Cloning and characterization of a novel δ12-fatty acid desaturase gene from the tree Sapium sebiferum. — Biotechnol. Lett, 29: 1573–6776, 2007.Google Scholar
- Sakuradani, E., Kobayashi, M., Ashikari, T., Shimizu, S.: Identification of δ12-fatty acid desaturase from arachidonic acid-producing Mortierella fungus by heterologous expression in the yeast Saccharomyces cerevisiae and the fungus Aspergillus oryzae. — Eur. J. Biochem. 261: 812–820, 1999.CrossRefPubMedGoogle Scholar
- Tao, F., Zhu, S.W., Fan, J.: Cloning and sequence analysis of maize FAD2 gene. — J. Plant Physiol. mol. Biol. 6: 649–656, 2006.Google Scholar
- Williams, J.P., Khan, M.U., Wong, D.: Fatty acid desaturation in monogalactosyl diacylglycerol of Brassica napus leaves during low temperature acclimation. — Plant Physiol. 6: 258–262, 1996.Google Scholar