Mouse nestin cDNA cloning and protein expression in the cytoskeleton of transfected cells.
Complementary DNA (cDNA) corresponding to mouse nestin intermediate filament protein, a specific marker for neural stem cells, was isolated and characterized. The complete sequence comprised 5983 base pairs encoding 1821 amino acids, and the deduced polypeptide was similar to rat (84%), hamster (73%), and human (62%) nestin. Southern blots showed that mouse nestin was a single-copy gene, and Northern blots detected a 6.0 kilobase mRNA transcript. When the cDNA was overexpressed as an enhanced green fluorescent fusion protein in COS7 cells, nestin immunoreactivity appeared in the filamentous cytoskeletal network. Accordingly, biologically active mouse nestin cDNA may offer an important new tool for stem cell research.
Yang
J
J
Institute of Biochemstry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, PR China.
Cheng
L
L
Yan
Y
Y
Bian
W
W
Tomooka
Y
Y
Shiurba
R
R
Jing
N
N
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D003001:Cloning, Molecular; D003599:Cytoskeleton / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins; D007381:Intermediate Filament Proteins / Q000737:chemistry / Q000235:genetics*; D008164:Luminescent Proteins / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins; D064231:Nestin; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D016415:Sequence Alignment; D014162:Transfection
D018076:DNA, Complementary; D007381:Intermediate Filament Proteins; D008164:Luminescent Proteins; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; C577486:Nes protein, rat; D064231:Nestin; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins