Gene Cloning and Identification of the Fatty Acid Biosynthetic Pathway in Camillia chekiangoleosa

Gene cloning and identification plays an important role in creating a transgenic plant that expresses certain genes conferring tolerance to abiotic stresses. Camellia chekiangoleosa is a native eastern Asian species that holds economic value in its production of tea-oil. The majority of research involved with this species is specific to planting/breeding. This study focuses to identify the molecular mechanisms of fatty acid synthesis that will yield the best quality and most efficiently made tea-oil. All the key genes involved with the fatty acid biosynthetic pathway of C. chekiangoleosa will be cloned and identified. For this particular study the following five were studied: stearoyl-ACP desaturase (SAD), acetyl-CoA carboxylase (ACC), biotin carboxylase (BC), 3-ketoacyl- CoA synthase (KCS), and acyl-acyl carrier protein thioesterase type B (FatB). A full-length cDNA sequence for each gene was amplified using PCR and cloned in E. coli by a pMD 19-T Vector (Takara Bio Inc., Japan). Sequencing was contracted out to BGI LifeTech Co. (Beijing, China). The BLAST analysis of each sequence identified species with highest similarities to C. chekiangoleosa based on protein sequences and structure. Once all the genes of interest are cloned and identified, the level of fatty acid synthesis at different developmental stages of C. chekiangoleosa seeds will be quantified. The overall objective of this study is to observe and measure the roles that each gene has in the production of fatty acids to produce a transgenic plant that will produce the best camellia oil.