Abstract
Oleoyl-ACP thioesterase (TE) is the chain length-determining enzyme in de novo biosynthesis of oleic acid. For cloning the gene encoding oleoyl-ACP TE from Brassica juncea, a PCR-amplified DNA probe was developed with the primers designed from the known sequences available in the Genbank. It was used to screen a genomic library of B. juncea constructed in λEMBL 3 to get the complete gene. A 4.0 kb Bam HI fragment from the clone X5.12 and hybridizing to the probe was sequenced. A stretch of 2606 by genomic sequence was found to contain an ORF of 1101 by with ∼ 0.4 kb untranslated regions at both the ends and 5 introns of varied lengths. By homology analysis, the ORF was found to encode an oleoyl-ACP thioesterase gene. The promoter region contains major cis elements required for transcription.
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