Cloning Of Fadx From Vernicia Fordii, Its Construction Of Antisense Expression Vector And Transformation Into Tobacco

Along with global fossil resources increasingly dried up, the development ofrenewable energy such as biodiese is imperative. Tung oil is one of good raw materialof biodiese which can be get easily and of good characteristics. But it should beimproved as other biodiese raw materials for that the main component of tung oil,a-eleostearic acid(18:3â–³^{9cis,11trans,13trans})is more than 90%. The tung oil has three doublebonds while the plant fatty acid one or two double bonds for high quality biodiese have.fadx is the key enzyme to convert linoleic acid into a-eleostearic acid. We try to clonethe fadx to change the composition of a-eleostearic acid by Antisense Expression.Total RNA were successfully extracted from developing tung seeds using improvedTRIzoL method, eDNA of fadx was cloned from Vernicia fordii by RT-PCR.Bioinformatic analysis showed that the cDNA was 1221bp long with anORF(14bp-1174bp) which encoded a polypeptide of 386 amino acids, and contains aspecific fatty acid desaturase structural domain. In order to introduce the gene intoplant, the cDNA was ligated to the pBI121 in the antisense orientation after digestedwith BamHI and SacI. The new vector named pBI121fadx was transformed to leaf oftobacco via Agrobacterium tumefaciens system.