Plasmid Constructs for Viral T Antigen Study

pCMV24-3xFLAG-Fbw7α wild-type and mutants, pCS2-5xMyc-Fbw7α, pCMV24-3xFLAG-Fbw1 wild-type and mutants, pCS2-HA-SV40 LT, and pCS2-HA-c-Myc have been described [21 (link), 23 (link), 24 (link), 27 (link), 29 (link), 31 (link), 32 , 42 (link)]. MCPyV T antigens were subcloned into pCS2, and/or pCS2-HA vectors using the In-fusion HD Cloning Kit (Clontech). The pCS2-MCPyV.57kT plasmid was generated by GENEWIZ TurboGENE Gene Synthesis. The MCPyV LT alanine scan mutagenesis was performed by ThermoFisher Scientific GeneArt. MCPyV T antigen mutants were generated using New England BioLabs Q5 Site-Directed Mutagenesis Kit, with primers designed using the NEBase Changer tool (New England Biolabs). pCGN.HA-Fbw7 and pCGN.HA-Fbw7 R465C were provided by Patrick Moore and Yuan Chang (University of Pittsburgh).

Free full text: Click here

Dye K.N., Welcker M., Clurman B.E., Roman A, & Galloway D.A. (2019). Merkel cell polyomavirus Tumor antigens expressed in Merkel cell carcinoma function independently of the ubiquitin ligases Fbw7 and β-TrCP. PLoS Pathogens, 15(1), e1007543.

Publication 2019

In-fusion HD Cloning Kit Q5 Site-Directed Mutagenesis Kit NEBase Changer tool

Alanine Mcpyv Mutagenesis Plasmid Primers Scan Site directed mutagenesis Sv40 Synthesis gene T antigen Vectors

Corresponding Organization : Seattle University

Top 5 similar protocols

Variable analysis

independent variables

PCMV24-3xFLAG-Fbw7α wild-type and mutants
PCMV24-3xFLAG-Fbw1 wild-type and mutants
MCPyV T antigen mutants

dependent variables

Not explicitly mentioned

control variables

PCS2-5xMyc-Fbw7α
PCS2-HA-SV40 LT
PCS2-HA-c-Myc
PCS2-MCPyV.57kT
PCGN.HA-Fbw7
PCGN.HA-Fbw7 R465C

controls

Positive control: pCS2-5xMyc-Fbw7α, pCS2-HA-SV40 LT, pCS2-HA-c-Myc
Negative control: pCGN.HA-Fbw7, pCGN.HA-Fbw7 R465C

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!