Title

Cloning of the Vacuolar H+-Pyrophosphatase Proton Pump Gene in Seashore Paspalum

School Name

Governor's School for Science and Mathematics

Grade Level

12th Grade

Presentation Topic

Botany

Presentation Type

Mentored

Abstract

There is a gene in some land plants that allows them to survive under higher salt stress than other plants can. One such species is the turf grass Seashore paspalum (Paspalum vaginatum), which contains a vacuolar H+ pyrophosphatase proton pump that allows the plant to more efficiently move sodium ions out of the water it is taking in. This allows it to live under conditions with higher salt stress. In this study, we isolated and amplified the gene that codes for this stronger proton pump. By utilizing polymerase chain reactions (PCRs) with varying primers and nested primers, we effectively amplified the selected target gene (PV26462). We then ligated this gene into a cloning vector and transferred it into E. coli, after which we isolated the recombinant plasmid DNA. We sent this isolated plasmid DNA to the New England BioLabs for sequencing to confirm that the gene had been properly inserted. In future research, blunt-end subcloning of the target gene will be performed. By using blunt-end subcloning, the gene will be introduced into an Agrobacterium. This product will be used to infect the calluses from samples of other turf grass species, allowing these transgenic plants to grow in higher-salt conditions.

Location

Neville 105

Start Date

4-14-2018 11:15 AM

Presentation Format

Oral and Written

COinS
 
Apr 14th, 11:15 AM

Cloning of the Vacuolar H+-Pyrophosphatase Proton Pump Gene in Seashore Paspalum

Neville 105

There is a gene in some land plants that allows them to survive under higher salt stress than other plants can. One such species is the turf grass Seashore paspalum (Paspalum vaginatum), which contains a vacuolar H+ pyrophosphatase proton pump that allows the plant to more efficiently move sodium ions out of the water it is taking in. This allows it to live under conditions with higher salt stress. In this study, we isolated and amplified the gene that codes for this stronger proton pump. By utilizing polymerase chain reactions (PCRs) with varying primers and nested primers, we effectively amplified the selected target gene (PV26462). We then ligated this gene into a cloning vector and transferred it into E. coli, after which we isolated the recombinant plasmid DNA. We sent this isolated plasmid DNA to the New England BioLabs for sequencing to confirm that the gene had been properly inserted. In future research, blunt-end subcloning of the target gene will be performed. By using blunt-end subcloning, the gene will be introduced into an Agrobacterium. This product will be used to infect the calluses from samples of other turf grass species, allowing these transgenic plants to grow in higher-salt conditions.