In this project, the maintenance of dynamic cycling of inorganic phosphate is discussed in terms of a phosphate transporter, ANTR3. Reverse genetics are applied with the help of bioinformatics and other web based tools which provided valuable initial information. The model organism Arabidopsis thaliana, both wild-type and mutants was cultivated hydroponically. Screening for homozygotes proceeded standardization of a protocol for isolation of plastids from the roots of these plants. Presence and absence of protein in root plastids was confirmed by western blotting in wild-type and mutants respectively. This was followed by functional and biochemical analysis of the protein by transport experiments using radioactively tagged phosphate. The development of a protocol for isolation of root plastids from roots of Arabidopsis thaliana with high yields has been successful. In-vitro studies by radioactive phosphate transport experiments were possible with the help of knock out mutants. It has been concluded that transport of Pi via ANTR3 is dependent either on an H+ or a Na+ gradient. Back exchange experiments have aided us in establishing its activity in export and import of Pi. Last but not the least, phenotypic analysis have observed larger biomass in mutants leading to the conclusion that even though other transporters are present the sink (roots) – source (leaves) balance is disturbed.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:liu-18938 |
| Date | January 2009 |
| Creators | Kuruvilla, Jacob |
| Publisher | Linköpings universitet, Institutionen för fysik, kemi och biologi |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Student thesis, info:eu-repo/semantics/masterThesis, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
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