In many organisms, trehalose protects against several environmental
stresses, such as heat, desiccation and salt, probably by stabilizing protein
structures and lipid membranes. Trehalose-6-phosphate synthase 1 (TPS1) is a
subunit of trehalose synthase complex in fungi / it plays a key role in the
biosynthesis of trehalose.
In this study, a TPS1 gene fragment in R. oryzae was cloned successfully
by PCR with primers designed according to eight hypothetical proteins found from
BLAST search which was performed by using S. cerevisiae TPS1 gene template.
The full length of R. oryzae TPS1 gene (designated RoTPS1) was attained by RTPCR
with primers specific to the 3& / #8242 / and 5& / #8242 / end of the RoTPS1 cDNA. The RoTPS1cDNA was composed of 2505 bps encoding a protein of 834 amino acids with a
molecular mass of 93.8 kDa. The amino acid sequence has relatively high
homology with the TPS1s of several other organisms.
RoTPS1 was further characterized by transformation into S. cerevisiae tps1
mutant. In galactose media, the growth curves of wild type, tps1 mutant and
transformant S. cerevisiae cells had a comparable pattern in general, tps1 mutant
reached to a higher maximum cell concentration compared to the others and wild
type had a slightly lower specific growth rate compared to the tps1 mutant and
transformed cells. Trehalose levels of transformant and wild type cells were
increased up to 37 mg/gdw in the stationary phase.
| Identifer | oai:union.ndltd.org:METU/oai:etd.lib.metu.edu.tr:http://etd.lib.metu.edu.tr/upload/3/12611025/index.pdf |
| Date | 01 September 2009 |
| Creators | Ozer Uyar, Gulsum Ebru |
| Contributors | Hamamci, Haluk |
| Publisher | METU |
| Source Sets | Middle East Technical Univ. |
| Language | English |
| Detected Language | English |
| Type | Ph.D. Thesis |
| Format | text/pdf |
| Rights | Access forbidden for 1 year |
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