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Journal of Biochemistry 2005 137(1):79-85; doi:10.1093/jb/mvi013
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© 2005 The Japanese Biochemical Society

CELL

Gene Trap Screening for Cell Surface and Extracellular Matrix Molecules Produced by Chondrocytes

Shin-ichi Shirasawa1,2, Masayuki Yoshimi1, Hideaki Kamochi1, Takeshi Muneta2, James H. Kimura3, Magnus Höök4 and Tamayuki Shinomura1,*

1 Tissue Regeneration and 2 Orthopedic Surgery, Department of Hard Tissue Engineering, Tokyo Medical and Dental University, Tokyo 113-8549; 3 Department of Surgery, John A. Burns School of Medicine, University of Hawaii at Manoa, Hawaii 96822, USA; and 4 Texas A&M Health Science Center, Institute of Biosciences and Technology, Texas 77030-3303, USA

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.

* To whom correspondence should be addressed. Tel: +81-3-5803-5450, Fax: +81-3-5803-0187, E-mail: t.shinomura.trg{at}tmd.ac.jp


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