Cloning of acyl-acyl carrier protein (ACP) thioesterase gene from Brassica juncea.
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 lambdaEMBL-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 lambda5.12 after a series of screening, sub-cloning and Southern hybridization.
Mandal
M N
MN
Division of Biochemistry, Indian Agricultural Research Institute, New Delhi 110012, India.
Santha
I M
IM
Lodha
M L
ML
Mehta
S L
SL
D001937:Brassica / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics; D015698:Genomic Library; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D018749:RNA, Plant / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D012639:Seeds / Q000201:enzymology; D013869:Thiolester Hydrolases / Q000235:genetics* / Q000378:metabolism*
D017931:DNA Primers; D018744:DNA, Plant; D018749:RNA, Plant; D011994:Recombinant Proteins; D013869:Thiolester Hydrolases; C018748:oleoyl-(acyl-carrier-protein) hydrolase