Acyl-acyl carrier protein (ACP) thioesterase is the chain-length-determining enzyme in de novo biosynthesis of plant fatty acids. For cloning the gene encoding acyl-ACP thioesterase from Brassica juncea genomic DNA was used as a template to amplify a 0.7 kb thioesterase fragment in a PCR with the primers designed from the known sequences available in the GenBank. This 0.7 kb fragment was used as a probe to study the expression of the gene in developing seeds and also to screen a genomic library of B. juncea constructed in λEMBL-3 to get the full length of the gene. A 4.0 kb BamHI fragment containing the full gene was finally cloned in a plasmid vector from a recombinant phage clone λ5.12 after a series of screening, sub-cloning and Southern hybridization.
Cloning of acyl-acyl carrier protein (ACP) thioesterase gene from Brassica juncea
- Views Icon Views
- Share Icon Share
M. N. A. Mandal, I. M. Santha, M. L. Lodha, S. L. Mehta; Cloning of acyl-acyl carrier protein (ACP) thioesterase gene from Brassica juncea. Biochem Soc Trans 1 December 2000; 28 (6): 967–969. doi: https://doi.org/10.1042/bst0280967
Download citation file: