关键词: 消减杂交, 基因, 克隆, 成骨细胞, 牵张力

Abstract:

bjective　In this experiment, genes sensitive to mechanical stretch in osteoblast like cells were cloned through
substractive 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 deoxyribornucleic 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 sub-
tractive cDNA library osteoblastic like cell stimulated with mechanical stretch was constructed through substractive 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 respec-
tively were identified to be novel, whichwere 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 substractive hybridization technique.

Key words: substractive hybridization, gene, clone, osteoblast, stretch