Production of Polyclonal Antibodies against Sucrose Transporter (SUT1) Protein Expressed in Escherichia coli BL21 and Application for Immunodiagnosis

Abstract

Sucrose transporter (SUT1) protein plays important roles in sucrose translocation from leaves to other organs in plants, therefore it is interested to study the existence of SUT1 protein in plants. Detection of SUT1 protein in plants can be done by using specific antibodies for that protein. This research was done to prepare SUT1 polyclonal antibodies by using SUT1 recombinant protein produced in Escherichia coli strain BL21. The production of SUT1 recombinant protein was done using fragment cDNA-SoSUT1 of sugarcane plants inserted inside plasmid pET28a, and expressed as a fusion protein containing N-terminal hexa-histidine tags. Expressed SUT1 was purified under denaturing conditions by affinity chromatography. Recombinant protein was purely used as antigen and was injected subcutanly in the back of female New Zealand White rabbits. Western Blot analysis using SUT1 polyclonal antibodies could detect the existence of SUT1 recombinant protein until the concentration 1.0 ng. Those antibodies could detect the existence of SUT1 protein at sugarcane plants.