The human PD-L1 promoter (position −831/+1)51 (link) was amplified by PCR from HCT116 genomic DNAs, and then cloned into the pGL3 basic reporter vector that contains firefly luciferase (Promega). The pRL-FL dicistronic reporter vector was generated from pcDNA3-RL-Polio IRES-FL vector21 (link) by deletion of Polio IRES. The full length and various deleted fragments of the PD-L1 5′-UTR were obtained by PCR using a HCT116 cDNA library, and then each was inserted between RL and FL of the pRL-FL vector. The phpRL-FL vector was generated by insertion of an inverted repeat sequence that forms a very stable RNA hairpin structure derived from phpBCL2-FL (Addgene) into the upstream RL of the pRL-FL vector. The Tagged-Cap and Tagged-LEF1 IRES plasmids were obtained from Addgene. The Tagged-PD-L1 IRES plasmid was constructed by replacing the LEF1 IRES with PD-L1 IRES from the Tagged-LEF1 IRES. The pLenti6.3-MS2-HB was generated by PCR using MS2-HB (Addgene) as a template to amplify MS2-HB, which was then cloned into the pLenti6.3 vector.21 (link) All sequences were verified by automated DNA sequencing.
Pgl3 basic reporter vector that contains firefly luciferase
Manufactured by Promega
The PGL3 basic reporter vector is a plasmid that contains the firefly luciferase gene. The luciferase enzyme catalyzes a bioluminescent reaction that generates light, which can be measured to quantify gene expression levels.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using pgl3 basic reporter vector that contains firefly luciferase
1
Characterization of PD-L1 Promoter and 5'-UTR
2
Characterization of PD-L1 Promoter and 5'-UTR
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The human PD-L1 promoter (position −831/+1)51 (link) was amplified by PCR from HCT116 genomic DNAs, and then cloned into the pGL3 basic reporter vector that contains firefly luciferase (Promega). The pRL-FL dicistronic reporter vector was generated from pcDNA3-RL-Polio IRES-FL vector21 (link) by deletion of Polio IRES. The full length and various deleted fragments of the PD-L1 5′-UTR were obtained by PCR using a HCT116 cDNA library, and then each was inserted between RL and FL of the pRL-FL vector. The phpRL-FL vector was generated by insertion of an inverted repeat sequence that forms a very stable RNA hairpin structure derived from phpBCL2-FL (Addgene) into the upstream RL of the pRL-FL vector. The Tagged-Cap and Tagged-LEF1 IRES plasmids were obtained from Addgene. The Tagged-PD-L1 IRES plasmid was constructed by replacing the LEF1 IRES with PD-L1 IRES from the Tagged-LEF1 IRES. The pLenti6.3-MS2-HB was generated by PCR using MS2-HB (Addgene) as a template to amplify MS2-HB, which was then cloned into the pLenti6.3 vector.21 (link) All sequences were verified by automated DNA sequencing.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!