pCMV-Luciferase (pCMV-Luc) was constructed previously (Kawakami et al., 2000 (link)). pZsGreen1-N1 and pDNAs of Mercury Pathway Profiling Luciferase system 1 (pTAL-Luc, pAP-1-Luc, and pNF-κB-Luc) were purchased from Clontech (Takara Bio Inc., Shiga, Japan). pCpG-free-LacZ and pCpG-free-Lucia were purchased from Invivogen (San Diego, CA). pCMV-Luc, pZsGreen1-N1, and pDNAs of Mercury Pathway Profiling Luciferase system 1 were amplified and purified as described previously (Kawakami et al., 2000 (link)). For amplification of the pCpG-free-plasmid (pCpG-free-LacZ and pCpG-free-Lucia), the E. coli competent strain GT115 (Invivogen) was used. Cy5-labeled pCMV-Luc and pZsGreen1-N1 were prepared using a Label IT Nucleic Acid Labeling Kit (Mirus Co., Madison, WI) according to the manufacturer’s protocol.
Pcpgfree lacz
Manufactured by InvivoGen
Sourced in United States
The PCpGfree-LacZ is a plasmid that contains the LacZ gene, which encodes the enzyme β-galactosidase. This plasmid lacks CpG motifs, making it suitable for studies involving gene expression in various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Pzsgreen1 n1, by Takara Bio (3 mentions)
Ptarget, by Promega (2 mentions)
Pcpgfree lucia, by InvivoGen (1 mentions)
Pnf κb luc, by Takara Bio (1 mentions)
Pap1 luc, by Takara Bio (1 mentions)
Ptal luc, by Takara Bio (1 mentions)
Endofree plasmid giga kit, by Qiagen (1 mentions)
Pcpgfree mcs vector, by InvivoGen (1 mentions)
5 protocols using pcpgfree lacz
1
Plasmid Construction and Labeling
2
Lentiviral LacZ Expression Cassette
3
Construction and Purification of Plasmids
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pCMV-Luc was constructed as previously described [20 (link)]. The hHGF expression vector without any CpG, pCpGfree-hHGF was custom-made by subcloning the CpG-depleted hHGF gene into the pCpGfree-mcs vector (InvivoGen, San Diego, CA, USA). pCMV-Luc and pZsGreen1-N1 (Takara Bio Inc., Shiga, Japan) were amplified in the Escherichia coli strain DH5α. pCpGfree-LacZ (InvivoGen, San Diego, CA, USA) and pCpGfree-hHGF were amplified in the E. coli strain GT115. Endofree Plasmid Giga Kit (QIAGEN GmbH, Hilden, Germany) was used to purified pDNA.
4
Cloning and Labeling Luciferase Plasmids
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pCMV-Luc, which contains a cytomegalovirus promoter and expresses firefly luciferase, was constructed as previously reported [18 (link)]. pCpG-free-LacZ was purchased from Invivogen (San Diego, CA, USA) and pZsGreen1-N1 was purchased from Clontech (Takara Bio Inc., Shiga, Japan). TMRhodamine-labeled pCMV-Luc was prepared using the Label IT-TM-Rhodamine Labeling Kit (Mirus Corp., Madison, WI, USA), according to the manufacturer’s instructions.
5
Lentiviral LacZ Expression Cassette
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The LacZ expression cassette consists of the EF1-α core promoter, the mouse cytomegalovirus enhancer, a synthetic intron, and the LacZ cDNA from the pCpGfree-LacZ (Invivogen, San Diego, CA, USA), and a SV40 late polyadenylation signal from pTarget™ (Promega, Madison, WI, USA).
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