Transformation of rapeseed [Brassica napus L.] plants with sense and antisense constructs of the fatty acid elongase gene

The biosynthetic pathways of saturated and unsaturated fatty acids consist of many steps controlled by various enzymes. One of the methods for improving oil quality is to change the fatty acid profile through genetic manipulation which requires isolation and characterization of the genes and other cis-acting elements, such as the promoter, involved in fatty acid biosynthesis. pketoacylCoA synthase [KCS] that is the key enzyme in erucic acid biosynthesis. This enzyme is involved in producing eicosanoeic [C20:1] and erucic acids [C22:1] from C18 fatty acids, and is encoded by the fatty acid elongase [FAE] gene. Specific primers were used to amplify the FAE gene and its promoter from genomic DNA by using PCR technique. The putative gene and its promoter were cloned in sense and antisense orientation into the plant expression vector [pBI121]. The sense and antisense constructs of the FAE gene were transformed via Agrobacterium-mediated transformation into low erucic acid rapeseed [LEAR such as PF] and high erucic acid cul-tivars [HEAR such as Maplus]. The transformed plants were screened on kanamycin-containing media and then analysed by PCR and Southern blotting techniques. Moreover, erucic acid content of the first generation of transgenic [T[0]] plants analysed with gas chromatography, showed significant changes in fatty acid composition of transgenic rapeseed plants containing sense and antisense constructs of the FAE gene

Cloning and expression of recombinant camelid single-domain antibody in tobacco

Antibodies provide a suitable tool in fundamental research and their high affinity and specificity make them invaluable for diagnostic and therapeutic applications. A promising alternative to conventional antibodies are the heavy chain antibodies [VHH] of Camelidae having short length, high solubility and stability are preferred to other antibody derivatives. In this study, our goal was production of recombinant VHH antibody fragments [against cancer associated mucin, MUC1] in tobacco plants. The VHH gene cDNA was cloned in TA vector and then subcloned into a plant expression binary vector pBI 121. The VHH gene was inserted into the plant genome by agrobacterium-mediated transformation. The presence of VHH gene in transformed plants was confirmed by PCR. Western blot analysis showed that the recombinant VHH protein was expressed in tobacco plant. ELISA results with MUC1 antigen confirmed that the biological activity and antigen-specific responses of the plant derived VHH protein compare favorably with that of the parent recombinant antibodies. This is the first report of production of camelied VHH antibody against tumor specific antigen from two-humped camel [Camelus bactrianus] in plants