Cloning The AccD Gene Coding Accase β-CT Of Gossypium Hirsutum Linn And Research On AccD Gene Overexpressing In Arabidopsis Thaliana

The fatty acids are essential components of the organism and in the common organisms, The fatty acids have a major physiological function. The fatty acids are the main ingredients of Membrane phospholipids, precursors of signal molecules and important energy;they have important value in industrial production and our daily life. Fatty acid biosynthesis is more complex,including Accase,FAS,FAD,FAE and so on. In recent years, people have taken research on the fatty acid biosynthesis, coded some related genes, stated fatty acids synthesis preliminarily, studied on regulation of fatty acids synthesis by genetic engineering. In this report, we study on fatty acid metabolism by accD gene overexpressing in plants, which coding Accaseβ-CT.The acetyl-CoA carboxylase (ACCase) is the rate-limiting enzyme of fatty acids synthesis, formate malonyl-CoA by Catalyticly. Malonyl-CoA is the important Substrate of fatty acid metabolism. ACCase include heteromeric and homomeric.β-CT of heteromeric Accase is coded by chloroplastgenome specially. We cloned accD and transit peptide of CAC3 gene from Gossypium hirsutum Linn and Arabidopsis thaliana. We constructed expression vector.The expression vector pBI121-CAC3tp-accD were transferred into Agrobacterium tumefaciens GV3101. Using infiltration and leaf discs method, the gene were transferred into Arabidopsis thaliana and tobacco cell. The seeds of Arabidopsis thaliana were selected on solid medium containing Kanamycin. Transferred leafs of tobacco were selected on solid medium containing Kanamycin. Transgenic Arabidopsis thaliana (T1) and tobacco plants were found containing purpose gene by PCR. We used RT-PCR methold to test the purpose gene transcription level. We tested the fatty acid content in leaves of transgenic Arabidopsis thaliana (T1), weight of thousands seeds of transgenic Arabidopsis thaliana (T1) and seed of every transgenic Arabidopsis thaliana (T1).This research not only revealed change in transgenic plant, but also provided a new way for fatty acid metabolism of Gossypium hirsutum Linn.