The LfKCS45 gene with a high sequence similarity to known 3-ketoacyl-CoA synthases of the mem- brane-bound fatty acid elongase was isolated from Lesquerella fendleri. The LfKCS45 gene has a 1464 bp open reading frame without introns, and is predicted to encode a polypeptide of 487 amino acids with an estimated molecular mass of 54.6 kD. High-stringency DNA blot analysis indicated that there were no closely related genes to LfKCS45 in the L. fendleri genome. Analysis of the fatty acid composition of transformed yeast revealed that expression of the LfKCS45 protein results in the synthesis of two novel very-long-chain fatty acids identiﬁed as C28:0 and C30:0. LfKCS45 was found to be not active with acyl-CoA substrates C16 to C24 in length. Reverse transcription-PCR experiments showed that the LfKCS45 gene is expressed only in L. fendleri root tips. Histochemical assays for GUS activity in Arabidopsis transformed with the LfKCS45 promoter-GUS fusion construct conﬁrmed this expression pattern and demonstrated that LfKCS45 transcription is restricted to the cells of the lateral root cap. Abbreviations: PCR, polymerase chain reaction; VLCFA, very-long-chain fatty acid, a fatty acyl group longer than 18 carbons; X:Y, a fatty acyl group containing X carbons with Y double bonds Introduction
Plant Molecular Biology – Springer Journals
Published: Apr 7, 2005
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