Expression of the chicken vimentin gene in transgenic mice: efficient assembly of the avian protein into the cytoskeleton.
To study expression and function of the vimentin gene, transgenic mice were generated by microinjecting the entire chicken gene plus 2.4 kilobases of 5' and 2.6 kilobases of 3' flanking sequences. All the transgenic mice obtained had incorporated multiple copies of the gene. RNA analyses demonstrated that the chicken vimentin gene was efficiently expressed in an appropriate tissue-specific pattern and that the transcripts were properly processed, as in chicken, giving rise to two RNAs. The vimentin transgene was predominantly expressed in lens at levels of up to 10-fold the endogenous level in every transgenic line studied. The chicken vimentin transcripts were efficiently translated into polypeptides that were modified posttranslationally and could assemble into the mouse cytoskeleton. Overexpression of the chicken vimentin gene did not obviously affect the expression of the endogenous gene at the RNA or the protein level. Immunofluorescence microscopy further demonstrated that the chicken protein was properly expressed spatially in lens. However, the levels were much higher in the transgenic animals.
Capetanaki
Y
Y
Department of Cell Biology, Baylor College of Medicine, Houston, TX 77030.
Starnes
S
S
Smith
S
S
D000818:Animals; D015152:Blotting, Northern; D002642:Chick Embryo; D002645:Chickens; D003599:Cytoskeleton / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005455:Fluorescent Antibody Technique; D005796:Genes; D007908:Lens, Crystalline / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D008845:Microinjections; D012016:Reference Values; D014746:Vimentin / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification
D014746:Vimentin