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.
Conclusion: It is an efficient approach to clone and study genes relative to mechanical stretch through subtractive hybridization technique.
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