a network of regulatory interactions governed by hfq

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