Expression of lincomycin biosynthetic gene cluster and its regulation
Project goals
The project aims in finding the organization of transcriptional units within the lincomycin biosynthetic cluster and identifying transcription starts. Based on previous results, e.g. the known sequence of the lincomycin biosynthetic gene cluster and partial knowledge of its pathway, putative regulatory genes were proposed. Functions of the proposed genes will be elucidated by inactivation and increasing the gene dose, respectively. The impact on lincomycin production will be observed. Preparation ofa cosmid library, gene inactivation, localization of transcription starts (primer extension), cDNA preparation and hybridization (microarray) will compose the methodical approach employed in the project. Determination of regulatory genes within the lincomycin biosynthetic cluster and their interference in expresion of individual genes and resulting antibiotic production are the expected results. Information on pathway-specific regulators and organization of transcription units is indispensable for
Keywords
Streptomyces lincolnensislincomycinpathway-specific regulators
Public support
Provider
Czech Science Foundation
Programme
Standard projects
Call for proposals
Standardní projekty 8 (SGA02005GA-ST)
Main participants
—
Contest type
VS - Public tender
Contract ID
204/05/0616
Alternative language
Project name in Czech
Exprese genů biosyntetického shluku linkomycinu a její regulace
Annotation in Czech
V navrhovaném projektu bude zjišťována organizace transkripčních jednotek a přesná místa počátků transkripce v biosyntetickém shluku linkomycinu. Na základě předchozích výsledků, tzn. znalosti uspořádání genů biosyntézy linkomycinu a částečné znalosti jeho biosyntetické dráhy, byly vytipovány možné regulační geny. Tyto geny budou jednak inaktivovány, jednak bude zvýšována jejich genová dose. Bude sledován vliv těchto zásahů na produkci linkomycinu. K základním metodám navrhovaného projektu patří příprava kosmidové knihovny, inaktivace genů, určení místa počátku transkripce (primer extension), příprava značené cDNA, a hybridizace (microarray). Očekávaným výsledkem projektu je určení regulačních genů linkomycinového biosyntetického shluku a zjištění, jakým způsobem ovlivňují expresi jednotlivých genů shluku a produkci antibiotika. Poznatky o dráhově specifických regulátorech a organizaci transkripčních jednotek umožní cílené manipulace biosyntetické dráhy linkomycinu.
Scientific branches
R&D category
ZV - Basic research
CEP classification - main branch
EB - Genetics and molecular biology
CEP - secondary branch
EE - Microbiology, virology
CEP - another secondary branch
EI - Biotechnology and bionics
10603 - Genetics and heredity (medical genetics to be 3)
10604 - Reproductive biology (medical aspects to be 3)
10605 - Developmental biology
10606 - Microbiology
10607 - Virology
10608 - Biochemistry and molecular biology
10609 - Biochemical research methods
20801 - Environmental biotechnology
20802 - Bioremediation, diagnostic biotechnologies (DNA chips and biosensing devices) in environmental management
20803 - Environmental biotechnology related ethics
20901 - Industrial biotechnology
20902 - Bioprocessing technologies (industrial processes relying on biological agents to drive the process) biocatalysis, fermentation
20903 - Bioproducts (products that are manufactured using biological material as feedstock) biomaterials, bioplastics, biofuels, bioderived bulk and fine chemicals, bio-derived novel materials
30101 - Human genetics
30401 - Health-related biotechnology
30402 - Technologies involving the manipulation of cells, tissues, organs or the whole organism (assisted reproduction)
30403 - Technologies involving identifying the functioning of DNA, proteins and enzymes and how they influence the onset of disease and maintenance of well-being (gene-based diagnostics and therapeutic interventions [pharmacogenomics, gene-based therapeutics])
30404 - Biomaterials (as related to medical implants, devices, sensors)
30405 - Medical biotechnology related ethics
40401 - Agricultural biotechnology and food biotechnology
40402 - GM technology (crops and livestock), livestock cloning, marker assisted selection, diagnostics (DNA chips and biosensing devices for the early/accurate detection of diseases) biomass feedstock production technologies, biopharming
40403 - Agricultural biotechnology related ethics
Completed project evaluation
Provider evaluation
U - Uspěl podle zadání (s publikovanými či patentovanými výsledky atd.)
Project results evaluation
HPLC methods for accurate determination of low quantities of products (lincomycin A and B) and intermediates (4-propyl-L-proline, methylthiolinkosamide, N-demethyl linkomycin and L-DOPA) of the lincomycin biosynthesis were elaborated. The methods were va
Solution timeline
Realization period - beginning
Jan 1, 2005
Realization period - end
Dec 31, 2007
Project status
U - Finished project
Latest support payment
May 2, 2007
Data delivery to CEP
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data delivery code
CEP08-GA0-GA-U/04:3
Data delivery date
Dec 16, 2008
Finance
Total approved costs
2,466 thou. CZK
Public financial support
2,466 thou. CZK
Other public sources
0 thou. CZK
Non public and foreign sources
0 thou. CZK
Basic information
Recognised costs
2 466 CZK thou.
Public support
2 466 CZK thou.
100%
Provider
Czech Science Foundation
CEP
EB - Genetics and molecular biology
Solution period
01. 01. 2005 - 31. 12. 2007