Cloning of a novel cDNA expressed during the early stages of fracture healing.
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.
Hadjiargyrou
M
M
Program in Biomedical Engineering, State University of New York at Stony Brook 11794-8181, USA. mhadjiargyro@ccmail.sunysb.edu
Halsey
M F
MF
Ahrens
W
W
Rightmire
E P
EP
McLeod
K J
KJ
Rubin
C T
CT
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002146:Bony Callus / Q000378:metabolism; D002454:Cell Differentiation; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics*; D017102:Fracture Healing / Q000235:genetics* / Q000502:physiology; D015870:Gene Expression; D066298:In Vitro Techniques; D007930:Leucine / Q000235:genetics* / Q000502:physiology; D008297:Male; D033681:Mitochondrial Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique; D051381:Rats; D017207:Rats, Sprague-Dawley; D014157:Transcription Factors / Q000235:genetics* / Q000502:physiology
D017931:DNA Primers; D018076:DNA, Complementary; D033681:Mitochondrial Membrane Transport Proteins; D000090622:Mitochondrial Precursor Protein Import Complex Proteins; D012333:RNA, Messenger; C114529:TIMM10B protein, human; C485424:Timm10b protein, rat; D014157:Transcription Factors; D007930:Leucine