期刊导航

论文摘要

用消减杂交技术克隆人成骨样细胞机械牵张力敏感基因的研究

Cloning Genes Sensitive to Mechanical Stretch in Osteoblasts through Substractive Hybridization Technique

作者:冯 雪1,丁 寅1,段银钟1,林 珠1,欧阳为明2,蒲 勤3

Author:FENGXue1, DING Yin1,DUAN Yin-zhong1,LIN Zhu1,OUYANG Wei-ming2,PU Qin3

收稿日期:2004-08-25          年卷(期)页码:2004,22(04):278-

期刊名称:华西口腔医学杂志

Journal Name:West China Journal of Stomatology

关键字:消减杂交,基因,克隆,成骨细胞,牵张力,

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

基金项目:国家自然科学基金资助项目(30170245,30370374)

中文摘要

目的 对人成骨样细胞机械牵张力敏感基因进行克隆研究。方法 对人成骨样细胞Saos-2进行二维方向上的加载,变形幅度为12%,牵拉频率为6周期/分钟,加载12 h后分别提取牵张力作用后Saos-2细胞及未作处理的正常Saos-2对照细胞的mRNA,反转录制备cDNA,将cDNA进行消减杂交,构建加载细胞与未加载细胞的差异表达基因的扣除cDNA文库,克隆后进行序列测定。结果 克隆到的基因片段中,功能已知基因片段15个,功能未知基因片段2个,其中牵张力敏感基因1(SSG1)位于9号染色体上,功能未知;牵张力敏感基因2(SSG2)位于18号染色体上,与转录因子2,4高度同源。结论 用消减杂交技术可以对人成骨细胞样机械牵张力敏感基因进行有效的克隆研究。

英文摘要

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.

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