Transformation of a Transposon Construct into Tomato for Functional Genomics Studies

Avirovik, Dragana

Transformation of a Transposon Construct into Tomato for Functional Genomics Studies

dc.contributor.author	Avirovik, Dragana	en
dc.contributor.committeechair	Pereira, Andy	en
dc.contributor.committeechair	Lawrence, Christopher B.	en
dc.contributor.committeemember	Tholl, Dorothea	en
dc.contributor.department	Biological Sciences	en
dc.date.accessioned	2014-01-17T09:00:08Z	en
dc.date.available	2014-01-17T09:00:08Z	en
dc.date.issued	2014-01-16	en
dc.description.abstract	Tomato (Solanum lycopersicum) is a member of the Solanaceae family. In this research project tomato, more specifically the M82 cultivar was chosen as a model plant for Agrobacterium-mediated gene transfer by cotyledon inoculation. Our objective was to transform tomato with a T-DNA construct bearing a transposon from maize that can be used for mutagenesis when it transposes or moves around the genome of the tomato. The vector used is a two-component in-cis Ac-Ds system which needs a single transformation event. It was proved that it worked in Arabidopsis and rice according to Trijatmiko (2005). The construct consists of the BAR gene conferring resistance to herbicide Basta, hygromycin (HYG) gene conferring resistance to the antibiotic hygromycin and the green fluorescent protein (GFP) gene, which are driven by specific plant promoters. The selectable marker genes such as HYG and BAR were used to select the rare transformation events by making the transformed tomato tissue resistant to the toxic chemicals (antibiotic and herbicide) compared to the untransformed tissue in which growth was inhibited. The results described consist of developing a transformation protocol which enabled the production of transgenic tomato lines by the help of the antibiotic augmetin (amoxicillin/clavulanic acid). The transgenic lines were tested through polymerase chain reaction (PCR) and herbicide bioassays.	en
dc.description.degree	Master of Science	en
dc.format.medium	ETD	en
dc.identifier.other	vt_gsexam:1611	en
dc.identifier.uri	http://hdl.handle.net/10919/24867	en
dc.publisher	Virginia Tech	en
dc.rights	In Copyright	en
dc.rights.uri	http://rightsstatements.org/vocab/InC/1.0/	en
dc.subject	Genetic transformation	en
dc.subject	functional genomics	en
dc.subject	expression of genes	en
dc.subject	Agrobacterium tumefaciens	en
dc.subject	Solanum lycopersicum.	en
dc.title	Transformation of a Transposon Construct into Tomato for Functional Genomics Studies	en
dc.type	Thesis	en
thesis.degree.discipline	Biological Sciences	en
thesis.degree.grantor	Virginia Polytechnic Institute and State University	en
thesis.degree.level	masters	en
thesis.degree.name	Master of Science	en