[Cloning genes sensitive to mechanical stretch in osteoblasts through subtractive hybridization technique].
OBJECTIVE
In this experiment, genes sensitive to mechanical stretch in osteoblast like cells were cloned through subtractive hybridization technique.


METHODS
Two dimensional mechanical stretch with deformation of 12% and frequency of 6 cycles was loaded on human osteoblastic like cell line Saos-2. Complementary deoxyribonucleic acid (cDNA) library of cells was constructed 12 h after loading, acting as tester. cDNA library of cells without loading was constructed, acting as driver. A subtractive cDNA library osteoblastic like cell stimulated with mechanical stretch was constructed through subtractive hybridization technique.


RESULTS
Of clones randomly selected from this library, fifteen genes were identified to be the differentially expressed genes. Comparing with the sequences published in GeneBank via Internet, two sequences located in chromosome 9 and 18 respectively were identified to be novel, which were named as stretch sensitive gene 1 and stretch sensitive gene 2.


CONCLUSIONS
It is an efficient approach to clone and study genes relative to mechanical stretch through subtractive hybridization technique.
Feng
Xue
X
Dept. of Orthodontics, Qindu Hospital, the Fourth Military Medical University, Xi'an 710032, China. cflfx@163.net
Ding
Yin
Y
Duan
Yin-zhong
YZ
Lin
Zhu
Z
Ouyang
Wei-ming
WM
Pu
Qin
Q
D002460:Cell Line; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D015723:Gene Library; D006801:Humans; D009693:Nucleic Acid Hybridization; D010006:Osteoblasts / Q000502:physiology*; D013314:Stress, Mechanical