Abstract
Using differential mRNA display (DD-PCR), a novel cDNA, FxC1 (Fracture Callus 1) was isolated from the early stages of a healing fractured femur. Utilizing 5' RACE PCR, a 598-bp full-length cDNA was obtained for FxC1 that contains an open reading frame (ORF) of 243 bp, encoding for an 80 amino acid protein. Within this ORF, a leucine zipper motif was present. In vitro transcription/translation of the full-length cDNA generated the expected 9-kDa protein. Northern analysis reveals that this gene is expressed in calluses harvested from post-fracture day 5, 7 and 10, as well as in several other tissues and bone-derived cell lines. During the differentiation of MC3T3 cells along the osteoblast lineage, FxC1 expression increases 3- to 4-fold during the production and deposition of matrix proteins, suggesting a possible role for this protein in cell differentiation.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Base Sequence
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Bony Callus / metabolism
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Cell Differentiation
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Cloning, Molecular
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DNA Primers / genetics
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DNA, Complementary / genetics*
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Fracture Healing / genetics*
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Fracture Healing / physiology
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Gene Expression
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In Vitro Techniques
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Leucine / genetics*
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Leucine / physiology
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Male
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Mitochondrial Membrane Transport Proteins
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Mitochondrial Precursor Protein Import Complex Proteins
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Molecular Sequence Data
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Open Reading Frames
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Protein Biosynthesis
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RNA, Messenger / genetics
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Random Amplified Polymorphic DNA Technique
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Rats
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Rats, Sprague-Dawley
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Transcription Factors / genetics*
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Transcription Factors / physiology
Substances
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DNA Primers
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DNA, Complementary
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Mitochondrial Membrane Transport Proteins
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Mitochondrial Precursor Protein Import Complex Proteins
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RNA, Messenger
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TIMM10B protein, human
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Timm10b protein, rat
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Transcription Factors
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Leucine