Molecular Characterization and Expression Analysis of long chain fatty acid acyl-CoA synthetase gene 1 (CoLACS1) from Camellia oleifera

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    long chain fatty acid acyl-CoA synthetase(LACS)plays an important role in fatty acid anabolism an catabolism. Based on seeds transcriptome Unigene sequence of Camellia oleifera named ‘Huashuo’(‘HS’), cDNA with complete sequence 2114 bp encoded LACS was isolated and cloned from Camellia oleifera seed, designated as CoLACS1 (GeneBank accession number:KJ960228). CoLACS1 had an open reading frame with 2088bp encoding 695 amino acids. sequence analysis demonstrated that the CoLACS1 possessed three blocks, the typical molecular characteristics of LACS, and belonged to LACS family. Phylogenetic analysis showed CoLACS1 is highly conserved and had the similarity( 78 %) with AtLACS7 and the similarity( over 80 % )with LACS6(peroxisomal)of Jatropha curcas, glycine max and Populus trichocarpa et al. The expression vector of pET30a-CoLACS1 was transformed to BL21 ( DE3) bacteria successfully,which was induced by 1 mmol/L IPTG to succeed in expressing the recombinant protein with molecular weight about 76 kD. The gene expression changes by qPCR,which were similar to those of RPKM in transcriptome data,indicate that the CoLACS1 expresses stably in different developmental stage of Camellia oleifera seeds and there was striking relationship between the gene expression of CoLACS1 and the accumulating of lipid in Camellia oleifera seed. The studies provide a theoretical basis for the futher reseach of gene regulating in lipid accumulating and metabolism of Camellia oleifera.

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  • Received:March 17,2016
  • Revised:May 26,2016
  • Adopted:July 01,2016
  • Online: January 17,2017
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