Integrated C. elegans Transgene Construction

The viral sensor construct and the gfp::sta-1 construct were generated by Gateway cloning using the MultiSite Gateway three-fragment vector construction kit (Life Technologies). Gateway entry clones containing each of the following were generated by standard techniques: sur-5 promoter, sdz-6 promoter, sta-1 coding sequence, eGFP(F64L/S65T) (eGFP stands for enhanced green fluorescent protein), and tbb-2 3′ untranslated region (3′UTR). The single-copy transgene was generated by transposase-mediated integration (MosSCI), as described previously (42 (link), 43 (link)), at insertion site ttTi5605 on chromosome II. Injection mixes contained 20 ng/µl of vector, 20 ng/µl of Mos1 transposase (only for MosSCI), and 5 ng/μl of a pharynx marker. Integration of the extrachromosomal array was performed by ethyl methanesulfonate (EMS) treatment (50 mM EMS for 4 h).

Free full text: Click here

Tanguy M., Véron L., Stempor P., Ahringer J., Sarkies P, & Miska E.A. (2017). An Alternative STAT Signaling Pathway Acts in Viral Immunity in Caenorhabditis elegans. mBio, 8(5), e00924-17.

Publication 2017

MultiSite Gateway three-fragment vector construction kit

3 utr Chromosome Clones Coding sequence Enhanced green fluorescent protein Ethyl methanesulfonate Mos1 transposase Pharynx Transgene Transposase Untranslated region Vector

Corresponding Organization :

Other organizations : Wellcome/Cancer Research UK Gurdon Institute, Wellcome Trust, University of Cambridge, Université Paris-Saclay, École Normale Supérieure Paris-Saclay, MRC London Institute of Medical Sciences, NIHR Imperial Biomedical Research Centre, Imperial College London

Top 5 similar protocols

Variable analysis

independent variables

Gateway cloning using the MultiSite Gateway three-fragment vector construction kit (Life Technologies)
Transposase-mediated integration (MosSCI) at insertion site ttTi5605 on chromosome II
Ethyl methanesulfonate (EMS) treatment (50 mM EMS for 4 h)

dependent variables

Integration of the extrachromosomal array

control variables

Injection mixes contained 20 ng/µl of vector, 20 ng/µl of Mos1 transposase (only for MosSCI), and 5 ng/μl of a pharynx marker

positive controls

None specified

negative controls

None specified

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!