Nucleic Acids Research, 1990, Vol. 18, No. 3 521-529
© 1990
MOLECULAR BIOLOGY |
Isolation of the chicken middle-molecular weight neurofilament (NF-M) gene and characterization of its promoter
ZMBH, Universität Heidelberg Im Neuenheimer Feld 282, D-6900 Heidelberg, FRG
* To whom correspondence should be addressed
Received October 13, 1989. Revised December 21, 1989. Accepted December 21, 1989.
We have isolated and sequenced genomic DNA clones covering the coding region of the chicken mid-size neurofilament (NF-M) gene and > 1 kb of its 5' upstream region. The NF-M gene contains two introns which both are located within the highly conserved C-terminal region of the rod domain. The 5' end of the corresponding mRNA was assigned to a G residue 40 nucleotides upstream of the translation start site and in appropriate distance from a potential TATA box. To functionally analyze the NF-M promoter, constructs carrying 112, 222, and 1026 nucleotides of the 5' upstream region in front of a luciferase reporter gene were tested for their capability to direct luciferase expression after transient transfection into various cell lines. Significant luciferase activity was recorded both in rat phaeochromocytoma (PC12) cells and murine fibroblasts. In PC12 cells, in which neurite outgrowth is induced by nerve growth factor (NGF), expression was stimulated up to 13-fold within 3 days of NGF treatment. This closely resembles expression of the endogenous NF-M gene in response to this hormone.
+Present addresses: Department of Biochemistry and Biophysics, University of California San Francisco, CA 94143, USA
Present addresses: Biologisches Institut, Universität Stuttgart, D-7000 Stuttgart 60, FRG and Institute for Molecular Biology, Bulgarian Academy of Science 1113 Sofia, Bulgaria
øZentrum für Molekulare Neurobiolgie UKE, Martinistr. 52, D-2000 Hamburg 20, FRG
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