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By Balch W.E., Channing J.D., Hall A.
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Extra info for Small GTPases and Their Regulators Part F
We also thank G w e n Mahon, Todd Palmby, and Ben Rushton for generation of expression constructs. D. D. C. (CA67771).  P r o t e i n T r a n s d u c t i o n : D e l i v e r y o f T a t - G T P a s e Proteins into Mammalian Cells Fusion B y ADAMINA VOCERO-AKBANI, M E E N A A . CHELLAIAH, KEITH A . HRUSKA, a n d STEVEN F. DOWDY Introduction Small GTPases, such as CDC42, Rac, and Rho, regulate the cytoskeletal architecture of the cell depending on the type of extracellular signals received. I-3 For instance, Rho plays an important role in the formation of actin stress fibers and focal adhesions.
Ecotropic viruses, regardless of whether they are produced in murine- or human-derived packaging lines, infect rodent cells. Amphotropic viruses can infect human cells in addition to rodent cells. Examples of packaging lines are Bosc23 cells,4 which produce ecotropic virus, and Phoenix 37 or Bing 4 cells, which produce amphotropic virus. Each of these cell lines is derived from 293T human embryonic kidney epithelial cells. We routinely infect NIH 3T3 and rat intestinal epithelial (RIE-1) cells with activated Ras family members [H-Ras(61L) and H-Ras (12V), KRas(12V), and N-Ras(12D) and N-Ras (61K)] (Table III) to establish stable cell lines.
This difference can be observed with GFP-tagged Ras protein in that, whereas GFP-N-Ras and GFP-H-Ras are visualized on rapidly motile peri-Golgi vesicles, GFP-K-Ras is not. This chapter details the methods used for tagging Ras proteins with GFP and imaging the fusion protein in live cells to observe subcellular localization and membrane trafficking. Expression Constructs The original clone of the green fluorescent protein (GFP) of the jellyfish Aequorea victoria isolated in 19927 has been widely disseminated and extensively mutated.
Small GTPases and Their Regulators Part F by Balch W.E., Channing J.D., Hall A.