[Transfection of articular chondrocytes with PcNDA3-hBMP3 and its stable expression].
OBJECTIVE
To explore the possibility of stable expression of PcDNA3-hBMP3 in cultured articular chondrocytes of rabbit.


METHODS
PcDNA3-hBMP3 was constructed using gene clone technique and recombined DNA technique. With the help of profectamine, the cultured articular chondrocytes were transfected with PcDNA3-hBMP3, and the evidence of successfully stable transfection in these cells could be obtained by positive northern blot.


RESULTS
The cultured articular chondrocytes of rabbits seemed to be polygonal, and its logarithmic growth phase was 2 - 4 days after cell inoculation. The two fragments cut from PcDNA3-hBMP3 by EcoR I and Xba I represented 5.4 kb and 1.4 kb by electrophoresis, which were confirmed to be the carrier and the fragment inserted originally, indicating that the construction of PcDNA3-hBMP3 was successful. The RNA extracted from cultured chondrocytes was screened for 4 weeks by G418 hibrided with the fragment cut from hBMP3 positively.


CONCLUSIONS
With the help of profectamine, the cultured articular chondrocytes can be transfected by recombined gene of PcDNA3-hBMP3 successfully, and their stable expression at 4 weeks after transfection is obtained.
Han
Y
Y
Department of Orthopedics, Xijing Hospital, Xi'an 710032.
Hu
Y
Y
Hui
H
H
Cui
D
D
Li
S
S
D000818:Animals; D055398:Bone Morphogenetic Protein 3; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis / Q000235:genetics*; D002358:Cartilage, Articular / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism*; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D004274:DNA, Recombinant / Q000096:biosynthesis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D055398:Bone Morphogenetic Protein 3; D019485:Bone Morphogenetic Proteins; D018076:DNA, Complementary; D004274:DNA, Recombinant; D012333:RNA, Messenger; D011994:Recombinant Proteins