Abstract

EXPRESSION OF RECOMBINANT HUMAN SERUM ALBUMIN IN NICOTIANA TABACUM

Pratheesh PT, Vineetha M and Muraleedhara Kurup G*

Abstract

Human serum albumin (HSA) is the most abundant protein in human blood plasma and also the most widely used protein by the pharmaceutical industry. Global demand for recombinant human serum albumin (rHSA) is increasing day by day due to the potential risks associated with plasma derived HSA and the limited availability of natural source. Though different expression systems are available for the production of rHSA, they all come up with a high price tag due to high cost of capitalization and production. Transgenic plants offer promising alternative for large scale, sustainable production of safe, functional, recombinant proteins of therapeutic and industrial importance. Therefore, as a preliminary step to produce human serum albumin in transgenic tobacco plants, HSA cDNA has been inserted into a plant expression vector pCAMBIA 1301 under the control of CaMV 35S promoter. The newly constructed vector pSBS.HSA was used to express HSA protein in Nicotiana tabacum using Agrobacterium mediated genetic transformation. Molecular confirmation of transformation was done through PCR analysis. Nucleotide sequencing was done to ensure that the 1.8 kb PCR fragment amplifying from transgenic tobacco is HSA itself. RT-PCR and SDS-PAGE confirmed that HSA is getting expressed in transgenic tobacco plants. Transgenic plants expressed recombinant HSA protein with a molecular mass of ∼66 kDa. Our results demonstrate feasibility for efficient expression of human serum albumin in transgenic tobacco plants using the newly constructed vector pSBS.HSA.

Keywords: Recombinant HSA. Nicotiana tabacum. Agrobacterium. Binary vector