Study of three proteins in uracil catabolism pathway
(2011) KEMP42 20102Department of Chemistry
- Abstract
- This study focused on expression, purification and functional characterization of three key proteins (Urc1p, Urc2p and Urc8p) in the newly discovered uracil catabolism (URC) pathway. A novel expression system was established by using Saccharomyces kluyveri as a host organism for expression of 6-His tagged Urc1p and Urc2p. Moreover, URC8 gene was cloned in pET151/D-TOPO vector and expressed in Escherichia coli. All three recombined proteins were purified on nickel columns. The SDS-PAGE showed Urc1p and Urc8p are of correct size and high purity. This novel expression system provides an alternative or a complementary choice for eukaryotic protein production. The functional characterization assay suggested that Urc1p might open the pyrimidine... (More)
- This study focused on expression, purification and functional characterization of three key proteins (Urc1p, Urc2p and Urc8p) in the newly discovered uracil catabolism (URC) pathway. A novel expression system was established by using Saccharomyces kluyveri as a host organism for expression of 6-His tagged Urc1p and Urc2p. Moreover, URC8 gene was cloned in pET151/D-TOPO vector and expressed in Escherichia coli. All three recombined proteins were purified on nickel columns. The SDS-PAGE showed Urc1p and Urc8p are of correct size and high purity. This novel expression system provides an alternative or a complementary choice for eukaryotic protein production. The functional characterization assay suggested that Urc1p might open the pyrimidine ring of uridine in the URC pathway and Urc8p is a NADPH dependent reductase converting malonic semialdehyde to 3-hydroxypropionate, which is one of the final products in the URC pathway. (Less)
Please use this url to cite or link to this publication:
http://lup.lub.lu.se/student-papers/record/3053636
- author
- Lv, You LU
- supervisor
-
- Jure Piskur LU
- organization
- course
- KEMP42 20102
- year
- 2011
- type
- H2 - Master's Degree (Two Years)
- subject
- keywords
- Uracil degradation, URC pathway, Saccharomyces kluyveri expression system, Proteinvetenskap
- language
- English
- id
- 3053636
- date added to LUP
- 2012-09-19 11:49:12
- date last changed
- 2012-09-19 11:49:12
@misc{3053636, abstract = {{This study focused on expression, purification and functional characterization of three key proteins (Urc1p, Urc2p and Urc8p) in the newly discovered uracil catabolism (URC) pathway. A novel expression system was established by using Saccharomyces kluyveri as a host organism for expression of 6-His tagged Urc1p and Urc2p. Moreover, URC8 gene was cloned in pET151/D-TOPO vector and expressed in Escherichia coli. All three recombined proteins were purified on nickel columns. The SDS-PAGE showed Urc1p and Urc8p are of correct size and high purity. This novel expression system provides an alternative or a complementary choice for eukaryotic protein production. The functional characterization assay suggested that Urc1p might open the pyrimidine ring of uridine in the URC pathway and Urc8p is a NADPH dependent reductase converting malonic semialdehyde to 3-hydroxypropionate, which is one of the final products in the URC pathway.}}, author = {{Lv, You}}, language = {{eng}}, note = {{Student Paper}}, title = {{Study of three proteins in uracil catabolism pathway}}, year = {{2011}}, }