MOESM3 of Metabolic pathway engineering using the central signal processor PII

Additional file 3: Figure S3. Cloning schemes for generation of the gene encoding PII(I86N) in Synechocystis sp. PCC 6803 showing all primer binding positions and restriction sites used in the construction. A) Spectinomycin resistance cassette (spec r) fused with a terminator sequence (Ter). B) PII-encoding gene glnB and downstream open reading frame slr0402. C) Engineered construct encoding PII(I86N) with spectinomycin resistance cassette inserted downstream of the variant glnB gene

The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds-3

column). The right column shows the corresponding bright field images of the transformed cells. The bars represent 25 μm. (B) Western blot analysis of protein extracts derived from transiently transformed tobacco leaves assayed for BiFC fluorescence before extraction (1–6). Immunodetection of the YFP-N fusion proteins (AHPs) was carried out with an antibody against c-myc-tag (anti c-myc) and of the YFP-C fusion protein (ARR22) with an antibody against the HA-tag (anti HA). M, protein marker.<p><b>Copyright information:</b></p><p>Taken from "The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds"</p><p>http://www.biomedcentral.com/1471-2229/8/77</p><p>BMC Plant Biology 2008;8():77-77.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2478664.</p><p></p

The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds-9

Ive RT-PCR. Data are expressed as mean +/- SD (n = 3) (B) Scheme of seed anatomy adapted according to Debeaujon and colleagues []. Note the location of the chalaza. (C) Confocal images of transgenic seeds at different developmental stages expressing a construct. Bright field, GFP images and overlays are presented from the left to the right. The bars represent 100 μm. (D) Immunolocalization of GFP on cryosections prepared from developing seeds of transgenic plants using a GFP-specific antibody (anti-GFP). The nuclei of the cells were visualised by DAPI staining (DAPI). The dotted line confines the chalaza area. The bars represent 20 μm.<p><b>Copyright information:</b></p><p>Taken from "The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds"</p><p>http://www.biomedcentral.com/1471-2229/8/77</p><p>BMC Plant Biology 2008;8():77-77.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2478664.</p><p></p

The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds-6

Ant and wild type (grey bars) by HPLC analysis as described in Methods. The data are expressed as mean ± SD (n = 5).<p><b>Copyright information:</b></p><p>Taken from "The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds"</p><p>http://www.biomedcentral.com/1471-2229/8/77</p><p>BMC Plant Biology 2008;8():77-77.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2478664.</p><p></p

The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds-0

Elay (drawn lines), which converges at the canonical AHPs (AHP1 to AHP5). Subsequently, the phosphoryl residues (P) are relayed to the A-type and B-type response regulator (ARRs), which are activiated by this phosphorylation and initiate the cellular responses (broken lines with arrow heads). Phosphorelay reactions catalyzed by receiver domains enable the phosphate flow in both directions. The phosphohistidine phosphatase ARR22 efficiently de-phosphorylates AHP2, 3 and 5, releases phosphate (P) and, therefore, acts a "sink" for phosphoryl residues. Especially in tissues, in which ARR22 is normally not expressed, this "sink" action strongly disturbs the phosphoload of the TCS network and interferes with TCS-regulated developmental processes (broken lines with vertical end line) causing dramatic phenotypic abnormalities. A similar "sink" function is reported for AHK4/CRE1 in the absence of cytokinin []. The pseudophosphotransfer protein AHP6 lacks the conserved His residue essential for phosphorelays and is proposed to inhibit the phosphate flow within the TCS network by competing with other AHPs for interactions with AHKs and ARRs [].<p><b>Copyright information:</b></p><p>Taken from "The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds"</p><p>http://www.biomedcentral.com/1471-2229/8/77</p><p>BMC Plant Biology 2008;8():77-77.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2478664.</p><p></p

The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds-4

On in black, UTR in grey), introns as thick lines, and the T-DNA insertions as triangles. The sequences at the insertion of the T-DNA (lower case letters) into the genome (upper case letters) are given for both alleles (, ). Arrows indicate the sites of primers used for RT-PCR analysis. (B) End-point RT-PCR analysis of the steady-state level of transcript. The cDNA was derived from total RNA extracted from siliques of the two allelic homozygous T-DNA insertion lines (, ) and wild type (Col-0). PCR was perfomed with the -specific primers indicated in (a) and, as a control, with -specific primers. To exclude any cross-contamination and contamination with genomic DNA, the RT-PCR was performed in the absence of total RNA or without its reverse transcription.<p><b>Copyright information:</b></p><p>Taken from "The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds"</p><p>http://www.biomedcentral.com/1471-2229/8/77</p><p>BMC Plant Biology 2008;8():77-77.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2478664.</p><p></p

The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds-7

Sphorylatable Asp74 (D74) was mutated either to a Glu () or an Asn () are shown.<p><b>Copyright information:</b></p><p>Taken from "The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds"</p><p>http://www.biomedcentral.com/1471-2229/8/77</p><p>BMC Plant Biology 2008;8():77-77.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2478664.</p><p></p

The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds-1

On channels for the RFP (ARR22:RFP) and GFP (marker fusion proteins) fluorescence and the overlay are indicated at the left. The lowest row shows the bright field images of the transformed cells. Marker genes are ARR2 for the nucleus, AHK5 for the cytoplasm and plasmalemma and GFP for nucleo-cytoplasmic distribution. The white arrows indicate the dense cytoplasm around the nucleus. The bars represent 10 μm.<p><b>Copyright information:</b></p><p>Taken from "The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds"</p><p>http://www.biomedcentral.com/1471-2229/8/77</p><p>BMC Plant Biology 2008;8():77-77.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2478664.</p><p></p

The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds-5

N bars) mutant and wild type (grey bars) by HPLC analysis as described in Methods. The data are expressed as mean ± SD (n = 5).<p><b>Copyright information:</b></p><p>Taken from "The response regulator ARR22 is a putative AHP phospho-histidine phosphatase expressed in the chalaza of developing seeds"</p><p>http://www.biomedcentral.com/1471-2229/8/77</p><p>BMC Plant Biology 2008;8():77-77.</p><p>Published online 15 Jul 2008</p><p>PMCID:PMC2478664.</p><p></p

De novo assembled transcriptome of Dandelion genotype A68

Transcriptome assembly based on RNAseq data of this study, deposited in NCBI sequence read archive, Bioproject number PRJNA316842. Trinity version trinityrnaseq-r2013-02-16