Screening of differentially expressed genes during adipocyte differentiation by suppression subtractive hybridization technique.
- Author:
Xiao-qing YI
1
;
Yan-feng XIAO
;
Chun-yan YIN
;
Er-di XU
Author Information
- Publication Type:Journal Article
- MeSH: Adipocytes; cytology; Cell Differentiation; genetics; Cell Line; Cloning, Molecular; Gene Expression Profiling; Gene Library; Genetic Vectors; Humans; Nucleic Acid Hybridization; methods; Transformation, Bacterial
- From: Journal of Zhejiang University. Medical sciences 2012;41(3):267-273
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo screening differentially expressed genes related to adipocyte differentiation.
METHODSTotal RNA extracted from the preadipocyte cell line SW872 was taken as the Driver and the total RNA from the differentiated adipocytes SW872 as the Tester. Suppression subtractive hybridization (SSH) was used to isolate the cDNA fragments of differentially expressed genes. The products of SSH were inserted into pGM-T vector to establish the subtractive library. The library was amplified through E.coli transformation and positive clones of the transformants were screened. Positive clones were sequenced. Nucleic acid similarity was subsequently analyzed by comparing with the data from GenBank.
RESULTSThere were 135 white clones in the cDNA library, 64 positive clones were chosen randomly and sequenced and similarity search revealed 34 genes which expressed differentially in adipocyte differentiation.
CONCLUSIONThe subtracted cDNA library for differentially expressed in adipocyte differentiation has been successfully constructed and the interesting candidate genes related to adipocyte differentiation have been identified.
