RNA polymerase II initiates from low complexity sequences so cells must reliably distinguish “real” from “cryptic” promoters and maintain fidelity to the former. Further, this must be performed under a range of conditions, including those found within inactive and highly transcribed regions. Here, we used genome-scale screening to identify those factors that regulate the use of a specific cryptic promoter and how this is influenced by the degree of transcription over the element.
We show that promoter fidelity is most reliant on histone gene transactivators (Spt10, Spt21) and H3-H4 chaperones (Asf1, HIR complex) from the replication-independent deposition pathway. Mutations of Rtt106 that abrogate its interactions with H3-H4 or dsDNA permit extensive cryptic transcription comparable with replication-independent deposition factor deletions. We propose that nucleosome shielding is the primary means to maintain promoter fidelity, and histone replacement is most efficiently mediated in yeast cells by a HIR/Asf1/H3-H4/Rtt106 pathway.
Physical association of Arabidopsis hypersensitive induced reaction proteins (HIRs) with the immune receptor RPS2.
Arabidopsis RPS2 is a typical nucleotide-binding leucine-rich repeat resistance protein, which indirectly recognizes the bacterial effector protein AvrRpt2 and thereby activates effector-triggered immunity (ETI). Previously, we identified two hypersensitive induced reaction (AtHIR) proteins, AtHIR1 (At1g09840) and AtHIR2 (At3g01290), as potential RPS2 complex components.
AtHIR proteins contain the stomatin/prohibitin/flotillin/HflK/C domain (also known as the prohibitin domain or band 7 domain). In this study, we confirmed that AtHIR1 and AtHIR2 form complexes with RPS2 in Arabidopsis and Nicotiana benthamiana using a pulldown assay and fluorescence resonance energy transfer (FRET) analysis.
Arabidopsis has four HIR family genes (AtHIR1-4). All AtHIR proteins could form homo- and hetero-oligomers in vivo and were enriched in membrane microdomains of the plasma membrane. The mRNA levels of all except AtHIR4 were significantly induced by microbe-associated molecular patterns, such as the bacterial flagellin fragment flg22.
Athir2-1 and Athir3-1 mutants allowed more growth of Pto DC3000 AvrRpt2, but not Pto DC3000, indicating that these mutations reduce RPS2-mediated ETI but do not affect basal resistance to the virulent strain. Overexpression of AtHIR1 and AtHIR2 reduced growth of Pto DC3000. Taken together, the results show that the AtHIR proteins are physically associated with RPS2, are localized in membrane microdomains, and quantitatively contribute to RPS2-mediated ETI.
Separation-of-function mutation in HPC2, a member of the HIR complex in S. cerevisiae, results in derepression of the histone genes but does not confer cryptic TATA phenotypes.
The HIR complex, which is comprised of the four proteins Hir1, Hir2, Hir3 and Hpc2, was first characterized as a repressor of three of the four histone gene loci in Saccharomyces cerevisiae. Using a bioinformatical approach, previous studies have identified a region of Hpc2 that is conserved in Schizosaccharomyces pombe and humans. Using a similar approach, we identified two additional domains, CDI and CDII, of the Hpc2 protein that are conserved among yeast species related to S. cerevisiae.
We showed that the N terminal CDI domain (spanning amino acids 63-79) is dispensable for HIR complex assembly, but plays an essential role in the repression of the histone genes by recruiting the HIR complex to the HIR-dependent histone gene loci. The second conserved domain, CDII (spanning amino acids 452-480), is required for the stability of the Hpc2 protein itself as well as for the assembly of the HIR complex.
In addition, we report a novel separation-of-function mutation within CDI of Hpc2, which causes derepression of the histone genes but does not confer other reported hir/hpc- phenotypes (such as Spt phenotypes, heterochromatin silencing defects and repression of cryptic promoters). This is the first direct demonstration that a separation-of-function mutation exists within the HIR complex.
The Saccharomyces cerevisiae histone chaperone Rtt106 mediates the cell cycle recruitment of SWI/SNF and RSC to the HIR-dependent histone genes.
In Saccharomyces cerevisiae, three out of the four histone gene pairs (HTA1-HTB1, HHT1-HHF1, and HHT2-HHF2) are regulated by the HIR co-repressor complex. The histone chaperone Rtt106 has recently been shown to be present at these histone gene loci throughout the cell cycle in a HIR- and Asf1-dependent manner and involved in their transcriptional repression.
Polyclonal HIRIP3 Antibody |
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| APR02968G | Leading Biology | 0.05ml | EUR 484 |
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Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human HIRIP3 . This antibody is tested and proven to work in the following applications: |
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HIRIP3 Polyclonal Antibody |
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| 30792-100ul | SAB | 100ul | EUR 252 |
HIRIP3 Polyclonal Antibody |
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| 30792-50ul | SAB | 50ul | EUR 187 |
anti- HIRIP3 antibody |
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| FNab03871 | FN Test | 100µg | EUR 585 |
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Description: Antibody raised against HIRIP3 |
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Anti-HIRIP3 antibody |
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| PAab03871 | Lifescience Market | 100 ug | EUR 412 |
HIRIP3 siRNA |
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| 20-abx919397 | Abbexa |
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HIRIP3 siRNA |
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| 20-abx919398 | Abbexa |
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anti-HIRIP3 |
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| YF-PA15655 | Abfrontier | 50 ul | EUR 363 |
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Description: Mouse polyclonal to HIRIP3 |
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anti-HIRIP3 |
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| YF-PA15656 | Abfrontier | 50 ug | EUR 363 |
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Description: Mouse polyclonal to HIRIP3 |
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pOTB7-HIRIP3 |
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| PVT18716 | Lifescience Market | 2 ug | EUR 231 |
Polyclonal HIRIP3 Antibody (Center) |
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| APR16639G | Leading Biology | 0.1ml | EUR 484 |
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Description: A polyclonal antibody raised in Rabbit that recognizes and binds to Human HIRIP3 (Center). This antibody is tested and proven to work in the following applications: |
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HIRIP3 Polyclonal Conjugated Antibody |
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| C31854 | SAB | 100ul | EUR 397 |
HIRIP3 Polyclonal Conjugated Antibody |
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| C30792 | SAB | 100ul | EUR 397 |
HIRIP3 Rabbit pAb |
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| A7014-100ul | Abclonal | 100 ul | EUR 308 |
HIRIP3 Rabbit pAb |
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| A7014-200ul | Abclonal | 200 ul | EUR 459 |
HIRIP3 Rabbit pAb |
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| A7014-20ul | Abclonal | 20 ul | EUR 183 |
HIRIP3 Rabbit pAb |
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| A7014-50ul | Abclonal | 50 ul | EUR 223 |
HIRIP3 cloning plasmid |
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| CSB-CL883612HU-10ug | Cusabio | 10ug | EUR 376 |
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Description: A cloning plasmid for the HIRIP3 gene. |
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Anti-HIRIP3 (3B3) |
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| YF-MA11069 | Abfrontier | 100 ug | EUR 363 |
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Description: Mouse monoclonal to HIRIP3 |
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HIRIP3 Blocking Peptide |
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| 33R-8235 | Fitzgerald | 100 ug | EUR 180 |
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Description: A synthetic peptide for use as a blocking control in assays to test for specificity of HIRIP3 antibody, catalog no. 70R-2185 |
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Mouse HIRIP3 shRNA Plasmid |
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| 20-abx982088 | Abbexa |
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Human HIRIP3 shRNA Plasmid |
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| 20-abx955558 | Abbexa |
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HIRIP3 ELISA KIT|Human |
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| EF010125 | Lifescience Market | 96 Tests | EUR 689 |
HIRIP3 Recombinant Protein (Rat) |
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| RP204605 | ABM | 100 ug | Ask for price |
HIRIP3 Recombinant Protein (Human) |
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| RP014686 | ABM | 100 ug | Ask for price |
HIRIP3 Recombinant Protein (Mouse) |
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| RP141530 | ABM | 100 ug | Ask for price |
HIRA-Interacting Protein 3 (HIRIP3) Antibody |
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| 20-abx113016 | Abbexa |
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HIRA-Interacting Protein 3 (HIRIP3) Antibody |
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| abx037953-100ug | Abbexa | 100 ug | EUR 391 |
HIRA-Interacting Protein 3 (HIRIP3) Antibody |
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| 20-abx005315 | Abbexa |
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HIRA-Interacting Protein 3 (HIRIP3) Antibody |
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| abx030421-400ul | Abbexa | 400 ul | EUR 523 |
HIRA-Interacting Protein 3 (HIRIP3) Antibody |
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| abx030421-80l | Abbexa | 80 µl | EUR 286 |
HIRA-Interacting Protein 3 (HIRIP3) Antibody |
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| abx233871-100ug | Abbexa | 100 ug | EUR 551 |
Hirip3 ORF Vector (Rat) (pORF) |
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| ORF068203 | ABM | 1.0 ug DNA | EUR 506 |
Hirip3 ORF Vector (Mouse) (pORF) |
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| ORF047178 | ABM | 1.0 ug DNA | EUR 506 |
HIRIP3 ORF Vector (Human) (pORF) |
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| ORF004896 | ABM | 1.0 ug DNA | EUR 95 |
HIRIP3 sgRNA CRISPR Lentivector set (Human) |
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| K0953201 | ABM | 3 x 1.0 ug | EUR 339 |
Hirip3 sgRNA CRISPR Lentivector set (Mouse) |
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| K4629701 | ABM | 3 x 1.0 ug | EUR 339 |
Hirip3 sgRNA CRISPR Lentivector set (Rat) |
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| K7156001 | ABM | 3 x 1.0 ug | EUR 339 |
HIRIP3 sgRNA CRISPR Lentivector (Human) (Target 1) |
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| K0953202 | ABM | 1.0 ug DNA | EUR 154 |
HIRIP3 sgRNA CRISPR Lentivector (Human) (Target 2) |
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| K0953203 | ABM | 1.0 ug DNA | EUR 154 |
HIRIP3 sgRNA CRISPR Lentivector (Human) (Target 3) |
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| K0953204 | ABM | 1.0 ug DNA | EUR 154 |
Hirip3 sgRNA CRISPR Lentivector (Mouse) (Target 1) |
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| K4629702 | ABM | 1.0 ug DNA | EUR 154 |
Hirip3 sgRNA CRISPR Lentivector (Mouse) (Target 2) |
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| K4629703 | ABM | 1.0 ug DNA | EUR 154 |
Hirip3 sgRNA CRISPR Lentivector (Mouse) (Target 3) |
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| K4629704 | ABM | 1.0 ug DNA | EUR 154 |
Hirip3 sgRNA CRISPR Lentivector (Rat) (Target 1) |
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| K7156002 | ABM | 1.0 ug DNA | EUR 154 |
Hirip3 sgRNA CRISPR Lentivector (Rat) (Target 2) |
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| K7156003 | ABM | 1.0 ug DNA | EUR 154 |
Hirip3 sgRNA CRISPR Lentivector (Rat) (Target 3) |
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| K7156004 | ABM | 1.0 ug DNA | EUR 154 |
Hirip3 3'UTR GFP Stable Cell Line |
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| TU255784 | ABM | 1.0 ml | Ask for price |
HIRIP3 3'UTR Luciferase Stable Cell Line |
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| TU009797 | ABM | 1.0 ml | EUR 1394 |
Hirip3 3'UTR GFP Stable Cell Line |
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| TU159519 | ABM | 1.0 ml | Ask for price |
Hirip3 3'UTR Luciferase Stable Cell Line |
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| TU205784 | ABM | 1.0 ml | Ask for price |
HIRIP3 3'UTR GFP Stable Cell Line |
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| TU059797 | ABM | 1.0 ml | EUR 1394 |
Hirip3 3'UTR Luciferase Stable Cell Line |
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| TU109519 | ABM | 1.0 ml | Ask for price |
HIRIP3 Protein Vector (Human) (pPB-C-His) |
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| PV019581 | ABM | 500 ng | EUR 329 |
HIRIP3 Protein Vector (Human) (pPB-N-His) |
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| PV019582 | ABM | 500 ng | EUR 329 |
HIRIP3 Protein Vector (Human) (pPM-C-HA) |
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| PV019583 | ABM | 500 ng | EUR 329 |
HIRIP3 Protein Vector (Human) (pPM-C-His) |
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| PV019584 | ABM | 500 ng | EUR 329 |
HIRIP3 Protein Vector (Rat) (pPB-C-His) |
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| PV272810 | ABM | 500 ng | EUR 603 |
HIRIP3 Protein Vector (Rat) (pPB-N-His) |
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| PV272811 | ABM | 500 ng | EUR 603 |
HIRIP3 Protein Vector (Rat) (pPM-C-HA) |
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| PV272812 | ABM | 500 ng | EUR 603 |
HIRIP3 Protein Vector (Rat) (pPM-C-His) |
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| PV272813 | ABM | 500 ng | EUR 603 |
HIRIP3 Protein Vector (Mouse) (pPB-C-His) |
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| PV188710 | ABM | 500 ng | EUR 603 |
HIRIP3 Protein Vector (Mouse) (pPB-N-His) |
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| PV188711 | ABM | 500 ng | EUR 603 |
HIRIP3 Protein Vector (Mouse) (pPM-C-HA) |
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| PV188712 | ABM | 500 ng | EUR 603 |
HIRIP3 Protein Vector (Mouse) (pPM-C-His) |
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| PV188713 | ABM | 500 ng | EUR 603 |
Human HIRA-Interacting Protein 3 (HIRIP3) ELISA Kit |
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| abx387793-96tests | Abbexa | 96 tests | EUR 911 |
Human HIRA- interacting protein 3, HIRIP3 ELISA KIT |
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| ELI-38603h | Lifescience Market | 96 Tests | EUR 824 |
HIRIP3 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV) |
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| LV627643 | ABM | 1.0 ug DNA | EUR 682 |
HIRIP3 Lentiviral Vector (Rat) (UbC) (pLenti-GIII-UbC) |
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| LV627647 | ABM | 1.0 ug DNA | EUR 682 |
HIRIP3 Lentiviral Vector (Rat) (EF1a) (pLenti-GIII-EF1a) |
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| LV627648 | ABM | 1.0 ug DNA | EUR 682 |
Mouse HIRA- interacting protein 3, Hirip3 ELISA KIT |
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| ELI-14456m | Lifescience Market | 96 Tests | EUR 865 |
HIRIP3 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Human) |
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| K0953205 | ABM | 3 x 1.0 ug | EUR 376 |
Hirip3 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Mouse) |
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| K4629705 | ABM | 3 x 1.0 ug | EUR 376 |
Hirip3 sgRNA CRISPR/Cas9 All-in-One Lentivector set (Rat) |
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| K7156005 | ABM | 3 x 1.0 ug | EUR 376 |
HIRIP3 sgRNA CRISPR/Cas9 All-in-One Lentivector (Human) (Target 1) |
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| K0953206 | ABM | 1.0 ug DNA | EUR 167 |
HIRIP3 sgRNA CRISPR/Cas9 All-in-One Lentivector (Human) (Target 2) |
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| K0953207 | ABM | 1.0 ug DNA | EUR 167 |
HIRIP3 sgRNA CRISPR/Cas9 All-in-One Lentivector (Human) (Target 3) |
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| K0953208 | ABM | 1.0 ug DNA | EUR 167 |
HIRIP3 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV-C-term-HA) |
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| LV627644 | ABM | 1.0 ug DNA | EUR 682 |
HIRIP3 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV-GFP-2A-Puro) |
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| LV627645 | ABM | 1.0 ug DNA | EUR 740 |
HIRIP3 Lentiviral Vector (Rat) (CMV) (pLenti-GIII-CMV-RFP-2A-Puro) |
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| LV627646 | ABM | 1.0 ug DNA | EUR 740 |
Hirip3 sgRNA CRISPR/Cas9 All-in-One Lentivector (Mouse) (Target 1) |
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| K4629706 | ABM | 1.0 ug DNA | EUR 167 |
Hirip3 sgRNA CRISPR/Cas9 All-in-One Lentivector (Mouse) (Target 2) |
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| K4629707 | ABM | 1.0 ug DNA | EUR 167 |
Hirip3 sgRNA CRISPR/Cas9 All-in-One Lentivector (Mouse) (Target 3) |
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| K4629708 | ABM | 1.0 ug DNA | EUR 167 |
Hirip3 sgRNA CRISPR/Cas9 All-in-One Lentivector (Rat) (Target 1) |
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| K7156006 | ABM | 1.0 ug DNA | EUR 167 |
Hirip3 sgRNA CRISPR/Cas9 All-in-One Lentivector (Rat) (Target 2) |
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| K7156007 | ABM | 1.0 ug DNA | EUR 167 |
Hirip3 sgRNA CRISPR/Cas9 All-in-One Lentivector (Rat) (Target 3) |
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| K7156008 | ABM | 1.0 ug DNA | EUR 167 |
H2B Antibody Antibody |
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| AF4659 | Affbiotech | 200ul | EUR 376 |
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Description: H2B Antibody Antibody detects endogenous levels of H2B. |
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CD11b Antibody Antibody |
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| ABD2911 | Lifescience Market | 100 ug | EUR 438 |
anti- Antibody^Polyclonal antibody control antibody |
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| LSMab09882 | Lifescience Market | 100 ug | EUR 438 |
Antibody |
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| A1360-500 | Biovision | Ask for price | |
Ly1 Antibody Reactive (LYAR) Antibody |
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| 20-abx123734 | Abbexa |
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Anti-Glycolipid Antibody (AGA) Antibody |
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| abx036399-100ug | Abbexa | 100 ug | EUR 391 |
The SWI/SNF and RSC chromatin remodeling complexes are both recruited to the HIR-dependent histone genes; SWI/SNF is required for their activation in S phase, whereas RSC is implicated in their repression outside of S phase. Even though their presence at the histone genes is dependent on the HIR complex, their specific recruitment has not been well characterized. In this study we focused on characterizing the role played by the histone chaperone Rtt106 in the cell cycle-dependent recruitment of SWI/SNF and RSC complexes to the histone genes.