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cDNA cloning and sequencing of MH2 domain of Smad2 from human dental pulp cells.

OBJECTIVE
The purpose of this study was to clone and sequence the cDNA of MH2 domain of Smad2 from human dental pulp cells.


METHODS
In this study, total RNA was isolated from primary cultured human dental pulp cells and reverse-transcribed into cDNA. The desired DNA product was obtained by nested PCR with 4 smad2 MH2 domain-specific primers. The segment was inserted into pBluescript II SK vector and the resulting plasmid was transformed into E. coli JM109. The double-strand cDNA of positive clone was sequenced by PE317-A automatic sequencing.


RESULTS
cDNA of MH2 domain of Smad2 was obtained from human dental pulp cells. The sequence was consistent with that cloned from a human kidney cDNA library. No mutation was found.


CONCLUSION
This study provides the first evidence of expression of smad2 in human dental pulp cells, and indicates that TGF-beta signaling may be mediated by Smad2 in human dental pulp cells. The cDNA cloned in pBluescript/S2MH2 could be used for further functional studies of Smad2 and MH2 domain in dental pulp cells.