pAcGFP1-Endo BioVector NTCC质粒载体菌种细胞基因保藏中心

pAcGFP1-Endo BioVector NTCC质粒载体菌种细胞基因保藏中心
pAcGFP1-Endo encodes a fusion protein consisting of the human RhoB GTPase containing an
N-terminal c-Myc epitope tag and the Aequorea coerulescens green fluorescent protein AcGFP1.
(Excitation maximum = 475 nm; emission maximum = 505 nm). The coding sequence of the AcGFP1
gene contains silent base changes, which correspond to human codon-usage preferences (1). SV40
polyadenylation signals downstream of the AcGFP1-c-Myc-RhoB fusion direct proper processing of
the 3' end of the mRNA. Sequences flanking AcGFP1 have been converted to a Kozak consensus
translation initiation site (2) to further increase the translation efficiency in eukaryotic cells. The
vector backbone also contains an SV40 origin for replication in mammalian cells expressing the
SV40 T-antigen. A neomycin resistance cassette (Neor), consisting of the SV40 early promoter, the
neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex
virus thymidine kinase (HSV TK) gene, allows stably transfected eukaryotic cells to be selected
using G418 (3). A bacterial promoter upstream of this cassette expresses kanamycin resistance in
E. coli. The pAcGFP1-Endo backbone also provides a pUC origin of replication for propagation in
E. coli and an f1 origin for single-stranded DNA production.
Use
The RhoB GTPase localizes the AcGFP1-c-Myc-RhoB fusion protein to the surface of vesicles of the
endocytic pathway (4) and allows the monitoring of intracellular membrane traffic during endocytosis.
For example these vesicles can be visualized in co-localization experiments such as receptor
internalization in living and fixed cells by using fluorescence microscopy. The c-Myc epitope allows
the monitoring of the fusion protein by Clontech Laboratories, Inc. c-Myc Monoclonal Antibody
(Cat. No. 631206) independently from AcGFP1 fluorescence. pAcGFP1-Endo can be transfected
into mammalian cells using any standard transfection method. If required, stable clones can be
selected using G418
BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
电话：+86-010-53513060
网址：www.biovector.net
【Supplier来源】BioVector NTCC Inc.
【Website网址】 http://www.biovector.net