Due to the incomplete JAZ sequences in the transcriptomic datasets, especially in the N and C termini of most lycophyte JAZ, we chose the longest JAZ sequence from H. myrsinites (CBAE2061511) and synthesized the corresponding coding sequence (cds SDS) (IDT technologies) from the codon encoding the first methionine upstream of the TIFY domain and including the attB tails. HmJAZ and AtJAZ3 chimera sequences were cloned into pKM596, pMpGW308, and pGADT7 (only for HmJAZ) via LR reaction (Invitrogen).
Pgadt7
Manufactured by Thermo Fisher Scientific
Sourced in United States
PGADT7 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is a thermal cycler designed for DNA amplification using the polymerase chain reaction (PCR) technique. The PGADT7 provides precise temperature control and programmable cycling for various PCR applications.
Automatically generated - may contain errors
Lab products found in correlation
Pgbkt7, by Thermo Fisher Scientific (2 mentions)
Pgex4t3, by Thermo Fisher Scientific (2 mentions)
Lr reaction, by Thermo Fisher Scientific (1 mentions)
Pentr d, by Thermo Fisher Scientific (1 mentions)
Anti ha peroxidase antibody, by Roche (1 mentions)
Pcr2.1 topo, by Thermo Fisher Scientific (1 mentions)
Petduet 1, by Thermo Fisher Scientific (1 mentions)
Pcrtm2.1 topo, by Thermo Fisher Scientific (1 mentions)
4 protocols using pgadt7
1
Synthesizing Lycophyte JAZ Sequences
2
Cloning and Transformation of Switchgrass NAP Genes
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The CDS of PvSSG PvNAP1 and PvNAP2 was amplified from the cDNA of a selected line "HR8" of the tetraploid switchgrass (P. virgatum L.) ecotype "Alamo" (Xu et al., 2011a) , cloned into the Gateway entry vector pENTR/D (Invitrogen Life Technologies, Carlsbad, CA, USA), and then subcloned into destination vectors, such as p2GWF7.0 for subcellular localization assay (Karimi et al., 2002) , pGBKT7 and pGADT7 (Invitrogen) for Y2H and transactivity assays, pVT1629 (Xu et al., 2011b) , for genetic transformation of switchgrass, and pEarlygate103 and pTA7001 for the transformation of Arabidopsis (Arabidopsis thaliana). A PvSSG-unique fragment was selected for RNAi (Supplemental Table S2) by generating a hairpin structure in the RNAi Entry vector, pGM-kannibal, and then subcloned the hairpin structure into the destination vector, pVT1629. The primers used for cloning and vector construction are shown in Supplemental Table S3.
3
Plasmid and Antibody Acquisition Protocol
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pCR2.1-TOPO, pETDuet-1, pGEX4T3, and pGADT7 plasmids were purchased from Invitrogen, Novagen, Life Sciences, and Clontech respectively. pCAM-BSD and pCAM-BSD-HA plasmids were kind gifts from Dr. C. Doerig (Monash University, Melbourne, Australia).
Monoclonal anti-HA, anti-penta His, anti-GST, anti-H3, and anti-HA peroxidase antibodies were purchased from Roche, Qiagen, Sigma-Aldrich, Millipore, and Abcam respectively. Anti-actin1 and anti-SOD1 antibodies were used as previously described (Daher et al., 2006 (link), 2010 (link)).
Monoclonal anti-HA, anti-penta His, anti-GST, anti-H3, and anti-HA peroxidase antibodies were purchased from Roche, Qiagen, Sigma-Aldrich, Millipore, and Abcam respectively. Anti-actin1 and anti-SOD1 antibodies were used as previously described (Daher et al., 2006 (link), 2010 (link)).
4
Reverse Genetic Studies in Plasmodium Species
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Plasmids pCRTM2.1-TOPO®, pQE30, pGEX4T3, pGADT7, and pGBKT7 were purchased from Invitrogen, Qiagen, Life Sciences and Clontech, respectively. Plasmids used in reverse genetic studies in P. falciparum, pCAM-BSD and pCAM-BSD-HA, were kind gifts from Prof. C. Doerig (Monash University, Melbourne, VIC, Australia). Plasmids used in P. berghei reverse genetic studies, p-TRAD4Ty-TetO7-HA-hDHFR and pBS-DHFR, were given by Prof. D. Soldati-Favre (University of Geneva, Switzerland) and Prof. R. Tewari (University of Nottingham, United Kingdom), respectively.
All primers used in this study are indicated in Supplementary Table1 .
All primers used in this study are indicated in Supplementary Table
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