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Pmturquoise2 h2a

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PmTurquoise2 H2A is a fluorescent protein construct that can be used to label histone H2A in live cells. It is derived from the Pontellina plumata turquoise fluorescent protein and can be used to visualize chromatin dynamics.

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Lab products found in correlation

Pdonr221, by Thermo Fisher Scientific (2 mentions) Bp cloning, by Thermo Fisher Scientific (2 mentions) Gateway system, by Thermo Fisher Scientific (1 mentions) Pmturquoise2 er, by Addgene (1 mentions) Pmturquoise2 mito, by Addgene (1 mentions) Pmturquoise2 peroxi, by Addgene (1 mentions) Pmturquoise2 golgi, by Addgene (1 mentions)

3 protocols using pmturquoise2 h2a

1

Plasmid Collection for Organelle Labeling

Cited 3 times
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These plasmids were a gift from Dorus Gadella: pmTurquoise2-Mito (Addgene plasmid #36208), pmTurquoise2-ER (Addgene plasmid #36204), pmTurquoise2-Peroxi (Addgene plasmid #36203), pmTurquoise2-Golgi (Addgene plasmid #36205) and pmTurquoise2-H2A (Addgene plasmid #36207)59 (link). pcDNA3.1-MCS-BirA-R118G-HA was a gift from Kyle Roux (Addgene plasmid # 36047)55 (link). RFP-Dcp1a and RFP-TIA1 were described before60 (link). MACROD1, MACROD2 and OARD1 (encoding TARG1) were amplified from Hela cDNA using primers with flanking attB sites suitable for the Gateway System (Invitrogen) and inserted into pDONR/zeo with a Gateway BP reaction. pcDNA5/FRT/TO-MACROD1, -MACROD2 or -TARG1 were generated by performing a Gateway LR reaction according to the manufacturer’s instructions. GFP-labelled plasmids were generated by LR reactions into pDEST47 and pDEST53. pcDNA3-mRuby2-MACROD1 and pcDNA3-mRuby2-MACROD1d77 were generated using primers with flanking HindIII and Nhel restriction sites for subsequent insertion into pcDNA3-mRuby2 for which the pcDNA5 constructs were used as PCR template. pcDNA3-BirA-R118G constructs with full length MACROD1, MACROD2 and TARG1 were generated using primers with flanking EcoRI and BamHI restriction sites and the pcDNA5 constructs as template.
Žaja R., Aydin G., Lippok B.E., Feederle R., Lüscher B, & Feijs K.L. (2020). Comparative analysis of MACROD1, MACROD2 and TARG1 expression, localisation and interactome. Scientific Reports, 10, 8286.
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2

Engineered mfap4 Promoter Driven Turquoise2

Cited 1 time
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The mfap4 promoter was PCR amplified using the primers 5′-CATGTTCTCGAGGCGTTTCTTGGTACAGCTGG-3′ and 5′-CATGTTGGATCCCACGATctaaagtcatgaagaaaga-3′. The product was subsequently cloned into p5E MCS using XhoI and BamHI sites. The native start codon was mutated using the primer 5′-CTGAGCTGTTGAGGAGAGAGTGAGAAG[ATT]GCAGTAAGTTCTGTGGCTGTTTTATTCC-3′ by inverse PCR with the backbone primers 5′-GTAAGTTCTGTGGCTGTTTTATTC-3′ and 5′-CTTCTCACTCTCTCCTCAACAG-3′. The final p5E mfap4 was then assembled by Gibson assembly using this single stranded oligo and the backbone.
To generate pME Turquoise2, we used the primers 5′-GGGGACAAGTTTGTACAAAAAAGCAGGCTggaccatggtgagcaagggcgaggag-3′ and 5′-GGGGACCACTTTGTACAAGAAAGCTGGGTttacttgtacagctcgtccat-3′ to amplify off pmTurquoise2 H2A (Addgene plasmid #3620235 (link)). The PCR product was subsequently cloned into pDONR221 by BP cloning (Invitrogen) to generate pME Turquoise2.
The mfap4:turquoise transgene construct was subsequently constructed by recombining p5E mfap4, pME Turquoise and p3E polyA into pDestTol2pA2 to generate pDestTol2; mfap4:turquoise.
Oehlers S.H., Cronan M.R., Scott N.R., Thomas M.I., Okuda K.S., Walton E.M., Beerman R.W., Crosier P.S, & Tobin D.M. (2014). Interception of host angiogenic signalling limits mycobacterial growth. Nature, 517(7536), 612-615.
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3

Engineered mfap4 Promoter Driven Turquoise2

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The mfap4 promoter was PCR amplified using the primers 5′-CATGTTCTCGAGGCGTTTCTTGGTACAGCTGG-3′ and 5′-CATGTTGGATCCCACGATctaaagtcatgaagaaaga-3′. The product was subsequently cloned into p5E MCS using XhoI and BamHI sites. The native start codon was mutated using the primer 5′-CTGAGCTGTTGAGGAGAGAGTGAGAAG[ATT]GCAGTAAGTTCTGTGGCTGTTTTATTCC-3′ by inverse PCR with the backbone primers 5′-GTAAGTTCTGTGGCTGTTTTATTC-3′ and 5′-CTTCTCACTCTCTCCTCAACAG-3′. The final p5E mfap4 was then assembled by Gibson assembly using this single stranded oligo and the backbone.
To generate pME Turquoise2, we used the primers 5′-GGGGACAAGTTTGTACAAAAAAGCAGGCTggaccatggtgagcaagggcgaggag-3′ and 5′-GGGGACCACTTTGTACAAGAAAGCTGGGTttacttgtacagctcgtccat-3′ to amplify off pmTurquoise2 H2A (Addgene plasmid #3620235 (link)). The PCR product was subsequently cloned into pDONR221 by BP cloning (Invitrogen) to generate pME Turquoise2.
The mfap4:turquoise transgene construct was subsequently constructed by recombining p5E mfap4, pME Turquoise and p3E polyA into pDestTol2pA2 to generate pDestTol2; mfap4:turquoise.
Oehlers S.H., Cronan M.R., Scott N.R., Thomas M.I., Okuda K.S., Walton E.M., Beerman R.W., Crosier P.S, & Tobin D.M. (2014). Interception of host angiogenic signalling limits mycobacterial growth. Nature, 517(7536), 612-615.
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