Transient expression of Aeromonas hydrophila HBNUAh01 outer membraneprotein A gene in tobacco leaf cells

Abstract:

Abstract:［Objective］To clone outer membrane protein A (ompA) gene from Aeromonas hydrophila HBNUAh01 and transiently express this protein in tobacco (Nicotiana tabacum) leaf cells． ［Methods］ A． hydrophila outer membrane protein A (AhompA) gene was amplified by PCR using A． hydrophila HBNUAh01 cells as template，and cloned into the pEASY-Blunt Simple vector for sequencing． After sequence confirmation，AhompA gene was inserted into expression vector pCAMBIA1300 containing a yellow fluorescent protein (YFP) gene，to obtain a recombinant expression plasmid． The recombinant expression plasmid was introduced into Agrobacterium tumefaciens GV3101 competent cells，and the positive clones were transfected to tobacco leaf cells． The expression of yellow fluorescent protein fused with AhompA was observed by Confocal Laser Scanning Microscope，and the mRNA of AhompA was detected by RT-PCR． ［Results］The AhompA gene cloned from A． hydrophila HBNUAh01 was 1032 bp． The fusion protein of AhompA and YFP was successfully expressed in tobacco leaf cells． ［Conclusion］The successful expression of the AhompA gene in tobacco leaf cells has laid a foundation for further investigation of prevention of limnobios diseases caused by A． hydrophila with plant vaccine．

Key words:Keywords: Aeromonas hydrophila，outer membrane protein，yellow fluorescent protein，tobacco，transient expression，plant vaccine