High molecular weight DNA purification instrument for metagenomics
High molecular weight DNA purification instrument for metagenomics
批准号:
7367898
负责人:
ANDRE MARZIALI
金额:
$20.58万
依托单位国家:
美国
项目类别:
财政年份:
2006
资助国家:
美国
项目状态:
已结题
起止时间:
2006-03-01 至 2009-08-31
关键词:
AddressAntibioticsBacterial DNABiochemical ReactionBiological MarkersBody FluidsBuffersCentrifugationCloningCompatibleComplexCoupledCytolysisDNADNA purificationDevelopmentElectrodesElectrophoresisEnvironmentEnzymesEscherichia coliFrequenciesFutureGelGene ClusterGenerationsGenomicsGrantInjection of therapeutic agentLaboratoriesLeftLengthLibrariesLicensingMalignant NeoplasmsMechanicsMetagenomicsMethodsMolecular WeightNatureNucleic AcidsOne-Step dentin bonding systemOrganismPathogen detectionPathway interactionsPerformancePharmaceutical PreparationsPolymerase Chain ReactionProceduresProcessProductionRNA DegradationRangeReagentRecoveryResearch PersonnelResourcesSamplingSiteSoilSolutionsSpeedStagingSystemTechniquesTechnologyTemperatureTestingVacuumWorkbacterial lysatebasecommercializationcostdesigndesireelectric fieldinhibitor/antagonistinstrumentinstrumentationinterestmacromoleculemicroorganismnext generationnovelprogramsprototypereconstructionsimulationsizesoil samplingtool
中文摘要
在自然界中观察到的所有微生物中,只有不到1%的微生物可以在实验室中培养,留下
英文摘要
Less than 1% of all microorganisms observed in nature can be cultured in the laboratory, leaving
researchers unable to study more than 99% of microorganisms in some environments - microorganisms that
sometimes have unique abilities such as synthesis of compounds that could find use as new drugs or
antibiotics. Metagenomics, the genomic reconstruction of unculturable microorganisms , is a powerful new
tool for accessing the untapped resources of these organisms. In one approach, large fragments ofDMA
from a sample containing unculturable microorganisms are extracted and cloned into a host such as E. coli
to produce a metagenomics library. The library is then screened for utility of expressed compounds.
Though new antibiotics and enzymes have been discovered with this method, successful production of
compounds, such as antibiotics, synthesized by the original microorganism presents the difficult challenge
of cloning fragments long enough to hold pathways encoded by gene clusters that are often over 100kb in
length. Existing purification techniques tend to shear genomic DMAto fragments of 50kb or less. The
discovery of a method to extract and purify high MW DMAfrom difficult samples such as soil will provide a
breakthrough for metagenomics that may enable the discovery of many future drugs and antibiotics.
We have recently demonstrated a non-linear electrophoretic method for DMA concentration that is
capable of DMA manipulation and concentration without mechanical handling (such as centrifugation or
pipetting). Extraction of DNA from bacterial lysate and 4,000 fold concentration factors have been
demonstrated. This method is an excellent candidate for the next generation of methods for DNA extraction
methods from complex samples such as soil or body fluids. We propose to develop an instrument to carry
out this concentration method, validating its performance on extraction of high molecular weight DNA on soil
samples for metagenomics studies aimed at discovery of new drugs and antibiotics. Licensing of this
technology will be pursued to address other compelling applications including DNA extraction from body
fluids for cancer biomarker and pathogen detection.
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