[Cloning genes sensitive to mechanical stretch in osteoblasts through subtractive hybridization technique]

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

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.

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.

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