Clathrin Light Chain A and Dynamin-2 Fluorescent Tagging

Human clathrin light chain A (CLTA) was amplified by PCR from a plasmid that was provided by L. Greene (NIH, USA)^{36 (link)}. This CLTA sequence is identical to the CLTA expressed in SK-MEL-2 cells. The African green monkey CLTA gene was amplified by reverse transcription PCR (RT–PCR) from poly-A mRNA of BSC-1 cells and subcloned into a pEGFP-N1 vector (Clontech) in which GFP was replaced with TagRFP-T (RFP)^{37 (link)}. TagRFP-T was a gift from R. Tsien (UC, San Diego, USA). A GTSGGS linker was placed between CLTA and RFP. Dynamin-2 (DNM2) was amplified by RT–PCR from poly-A mRNA of SK-MEL-2 cells and subcloned into a pEGFP-N1 vector with a GTSGGS linker between DNM2 and GFP.

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Doyon J.B., Zeitler B., Cheng J., Cheng A.T., Cherone J.M., Santiago Y., Lee A.H., Vo T.D., Doyon Y., Miller J.C., Paschon D.E., Zhang L., Rebar E.J., Gregory P.D., Urnov F.D, & Drubin D.G. (2011). Rapid and efficient clathrin-mediated endocytosis revealed in genome-edited mammalian cells. Nature cell biology, 13(3), 331-337.

Publication 2011

African green monkey Cells Clathrin light chain Dynamin 2 Gene Human Plasmid Poly a mrna Reverse transcription Vector

Corresponding Organization :

Other organizations : University of California, Berkeley, Sangamo BioSciences (United States)

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Variable analysis

independent variables

Plasmid provided by L. Greene (NIH, USA) containing human clathrin light chain A (CLTA) sequence
African green monkey CLTA gene amplified from poly-A mRNA of BSC-1 cells
Dynamin-2 (DNM2) amplified from poly-A mRNA of SK-MEL-2 cells

dependent variables

Not explicitly mentioned

control variables

SK-MEL-2 cell line as the source of CLTA sequence
BSC-1 cell line as the source of African green monkey CLTA gene
SK-MEL-2 cell line as the source of Dynamin-2 (DNM2)

positive controls

Not mentioned

negative controls

Not mentioned

Annotations

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