Hevea genetic transformation for enhanced recombinant pharmaceutical production by the use of hevein promoter

Pharmaceuticals produced in the latex cytosol of genetically transformed Hevea can be harvested nondestructively by conventional tapping. This study evaluates the promoter activity of the 5’-untranslated upstream regulating region of hevein gene, which encodes the most abundant soluble protein in Hevea latex. Constructs were prepared with the test hevein promoter fragments Hev P1 (0.35kb), Hev P2 (0.45kb) and Hev P3 (0.73kb) that were inserted 5’ to the pharmaceutical genes i.e. human protamine 1 (HP1) and human atrial natriur tic factor (HANF), in pGPTV-Kan expression vector. The expression vectors containing HP1 and HANF were electroporated into Agrobacterium tumefaciens GV2260 containing supervirulent plasmid pToK47, which were then used to infect Hevea anther callus. The growth of the putative transformed Hevea callus was monitored on kanamycin selection media. The presence of the pharmaceutical genes (HP1 and HANF), the hevein promoter fragments, and nptII selection marker were verified by PCR on sampled putative transformed callus. The remaining callus tissues will be sub-cultured and transferred into differentiation media for embryoids formation and plantlet regeneration

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Abstract. -Objectives: Hevea brasiliensis extract could potentially be employed as a relatively low cost resource for various anti-fungal activities due to the simplicity of latex preparation and the abundance of latex that can be obtained in rubber producing regions. The present study was aimed at examining the species specific anti-fungal property of H. brasilensis latex C-serum against Aspergillus niger. Results: The results showed that the latex Cserum exerted a specific antifungal activity against Aspergillus niger, but not Candida albicans. Low toxicity of the C-serum was demonstrated in Brine Shrimp Lethality Test (BSLT) with an LC 50 value of 98.4 mg/ml. Conclusions: Pending further more elaborated investigations, H. brasiliensis latex C-serum, with its species specific anti-fungal and cancer-origin cell line specific anti-proliferation properties, would probably contribute in healthcare in addition to its traditional role in polymer industry