Cloning of a novel cDNA expressed during the early stages of fracture healing

Biochem Biophys Res Commun. 1998 Aug 28;249(3):879-84. doi: 10.1006/bbrc.1998.9167.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Bony Callus / metabolism
  • Cell Differentiation
  • Cloning, Molecular
  • DNA Primers / genetics
  • DNA, Complementary / genetics*
  • Fracture Healing / genetics*
  • Fracture Healing / physiology
  • Gene Expression
  • In Vitro Techniques
  • Leucine / genetics*
  • Leucine / physiology
  • Male
  • Mitochondrial Membrane Transport Proteins
  • Mitochondrial Precursor Protein Import Complex Proteins
  • Molecular Sequence Data
  • Open Reading Frames
  • Protein Biosynthesis
  • RNA, Messenger / genetics
  • Random Amplified Polymorphic DNA Technique
  • Rats
  • Rats, Sprague-Dawley
  • Transcription Factors / genetics*
  • Transcription Factors / physiology

Substances

  • DNA Primers
  • DNA, Complementary
  • Mitochondrial Membrane Transport Proteins
  • Mitochondrial Precursor Protein Import Complex Proteins
  • RNA, Messenger
  • TIMM10B protein, human
  • Timm10b protein, rat
  • Transcription Factors
  • Leucine

Associated data

  • GENBANK/AF061242