Algae. 2013; 28(4): 379-387.
Expression of bovine lactoferrin N-lobe by the green alga, Chlorella vulgaris
Jungmo Koo1,*, Dongjun Park1 and Hakeung Kim1
1GENOBIO Co., Daejeon 306-230, Korea
ABSTRACT
The purpose of this study was to express bovine lactoferrin N-lobe in Chlorella vulgaris, a green microalga, using the pCAMBIA1304 vector. Chlorella-codon-optimized bovine lactoferrin N-lobe (Lfb-N gene) was cloned in the expression vector pCAMBIA1304, creating the plasmid pCAMLfb-N. pCAMLfb-N was then introduced into C. vulgaris by electrotransformation. Transformants were separated from BG-11 plates containing 20 μg mL-1 hygromycin. Polymerase chain reaction was used to screen transformants harboring Lfb-N gene. Finally, total soluble protein was extracted from the transformants, and the expression of Lfb-N protein was detected using western blotting. Using this method, we successfully expressed bovine lactoferrin in C. vulgaris. Therefore, our results suggested that recombinant lactoferrin N-lobe, which has many uses in the biomedical and pharmaceutical industries, can be produced economically.
Keywords :
Chlorella vulgaris; electrotransfomation; expression; hygromycin; lactoferrin