a network of regulatory interactions governed by hfq
Post on 19-Jan-2016
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A network of regulatory interactions governed by Hfq.Hfq has been shown to bind each of the ncRNA (circled) and mRNAs (outer ring) facilitatingthe post-transcriptional gene regulation that responds to the indicated environmental stress.
From the Feig LaboratoryWayne State University WEB Page
From the Feig LaboratoryWayne State University WEB Page
Secondary structures of DsrA (top) and rpoS mRNA 5'-UTR(bottom) The color coding of DsrA indicates the accepted domain structure. For rpoS, the open reading fram is shown in green and the region complementary to DsrA (bases 11 - 32 in DsrA) is shown in lavender.
Generic model through which regulatory ncRNAs can relieve translational repression of a complementary mRNA through strand displacement. Equivalent models can be used to explain translational repression as well.
Principle of pyrosequencing
http://www.s3.kth.se/mst/research/projects/beadbasedmicrofluidic/index.shtml
How to detect the gene expression? (1) Northern Blot analysis
(2) Primer extension analysis (3) S1 nuclease mapping
(4) Nuclease protection assay
(5) RT-PCR
(6) Large-scale cDNA sequencing
(7) Microarray analysis
(8) 5’-, 3’- RACE (Rapid Amplification of cDNA En
ds)(9) Reporter gene assay
Northern Blot Hybridization
Figure 4.12. Southern hybridization. (A) Transfer of DNA from the gel to the membrane. (B) The membrane is probed with a radioactively labeled DNA molecule. On the resulting autoradiograph, one hybridizing band is seen in lane 2, and two in lane 3.
Primer Extension Analysis
S1 Nuclease Mapping Analysis
Gene Gene product Assay
lacZ β-galactosidase Histochemical test
uidA β-glucuronidase Histochemical test
lux Luciferase Bioluminescence
GFP Green fluorescent protein
Fluorescence
Reporter Gene Assay
Luciferase gene
Luciferase Activity (Promega)
PITX2-LACZ Staining of mouse embryo day 11
PITX2: bicoid-related transcription factor
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