الفهرس | Only 14 pages are availabe for public view |
Abstract Thaumatin-like proteins (TLPs) constitute a homogeneous family, members of which are produced by plants in response to different kinds of stress. NP24 is one such salt-induced protein from tomato which is a rich source of the protein, and belongs to this family. NP24 is 24 kDa ( 207-amino-acid). Antimicrobial thaumatin-like protein (TLP) found in tomato fruits. One isoform (NP24I) of the thaumatin-like protein NP24 has been expressed mainly in the outer pericarp of tomato fruit and is reported to play a possible role in ripening of the fruit in addition to its antimicrobial activity. In this study, the total RNA was successfully isolated from the outer pericarp of ripe (red) tomato fruit. cDNA was prepared and amplified using the suitable PCR programe .The gene was cloned into The Mach1™- T1® E. coli cells, Then subcloned into the overexpression vector pET-28a (+) using BL21 expression bacteria. The SDS-PAGE analysis showed that the gene was overexpressed in E. coli as inclusion bodies. Optimization of the recombinant NP24I protein solubility was achieved by decreasing both expression temperature and IPTG concenteration (cold induction). The recombinant NP24I protein was purified using Ni-NTA resin. The antimicrobial activity of the recombinant NP24I protein was tested. |