Abstract:
Brazzein- a low-calorie natural sweetener is an effective substitute for deleterious effects of sucrose. Uptill now, limited
production of recombinant brazzein protein using heterologous expression system along with solubility and low yield
challenges are reported. Foreseeing the potential of brazzein gene in therapeutics, molecular strategies are required to
optimize production of recombinant brazzein using different prokaryotic and eukaryotic expression systems. Therefore,
in this study brazzein gene synthesized from Bio Basic Inc. (BBI) which was provided in pUC57 cloning vector. The
gene was retrieved from cloning vector by performing PCR and cloned in plant expression vector pBI121 with
CaMV35S promoter region. Heat shock or calcium chloride method was optimized for transformation into E. coli (DH5-
alpha) with recombinant vector pBI121. Kanamycin resistence selection, colony PCR and transformation efficiency
analysis were performed to analyze the successful cloning procedure. Transformed cultures having cloned brazzein gene
in pBI121 vector with CaMV35S promoter could be used to transform different plants such as, sorghum, sugarcane and
turnip for sweet taste enhancement in future and may provide plateform for commercial scale production of therapeutic
proteins using crop plants as bioreactor.
