CONSTRUCTION OF THE OseIF4AIIa-GFP EXPRESSION VECTOR AND GENERATION OF RECOMBINANT Agrobacterium tumefaciens FOR FUNCTIONAL ANALYSIS OF THE GENE IN RICE

Abstract

This study aimed to isolate and construct an expression vector carrying the OseIF4AIIa gene from rice (Oryza sativa) to support subsequent functional analyses. The gene is hypothesized to encode a DEAD-box RNA helicase involved in the plant's response to adverse environmental conditions. The coding sequence of OseIF4AIIa was amplified from rice cDNA using polymerase chain reaction, producing a fragment of 1,261 bp. The amplified fragment was purified, cloned into the yT&A cloning vector, and then subsequently transferred into the binary vector pMDC85 to generate the fluorescent fusion construct pMDC85-OseIF4AIIa-GFP. The recombinant vector was successfully introduced into Agrobacterium tumefaciens via electroporation and confirmed by polymerase chain reaction. The entire process, from gene isolation to expression vector construction, resulted in a recombinant bacterial strain that carries the target gene construct. This study provides a technical foundation for future rice transformation experiments and functional analysis of OseIF4AIIa.