Gene trap screening for cell surface and extracellular matrix molecules produced by chondrocytes.
We have developed a simple and unique strategy for identifying cell surface and extracellular matrix molecules produced by chondrocytes. Our strategy comprises two methods, retrovirus-based signal sequence gene trapping and culturing of Swarm rat chondrosarcoma chondrocytes. After infection with a retrovirus vector and isolation of hygromycin-resistant clones, trapped genes could be easily identified by the 5' rapid amplification of cDNA ends (5' RACE) method. Furthermore, the treatment of isolated clones with gadolinium chloride enabled us to determine whether the trapped gene expression was dependent on the state of chondrogenic differentiation.
Shirasawa
Shin-ichi
S
Tissue Regeneration and Orthopedic Surgery, Department of Hard Tissue Engineering, Tokyo Medical and Dental University, Tokyo 113-8549, Japan.
Yoshimi
Masayuki
M
Kamochi
Hideaki
H
Muneta
Takeshi
T
Kimura
James H
JH
Höök
Magnus
M
Shinomura
Tamayuki
T
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D019902:Chondrocytes / Q000502:physiology*; D003001:Cloning, Molecular / Q000379:methods*; D005109:Extracellular Matrix / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D051381:Rats
D008565:Membrane Proteins