Cloning differentially regulated genes from chondrocytes using agarose gel differential display

被引:22
作者
Jefferies, D [1 ]
Botman, M [1 ]
Farquharson, C [1 ]
Lester, D [1 ]
Whitehead, CC [1 ]
Thorp, BH [1 ]
Houston, B [1 ]
机构
[1] Roslin Inst, Roslin EH25 9PS, Midlothian, Scotland
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION | 1998年 / 1396卷 / 03期
关键词
agarose gel differential display; chondrocyte hypertrophy; cell culture; gene expression;
D O I
10.1016/S0167-4781(97)00234-0
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The technique of RNA differential display has been used extensively to clone differentially expressed genes from a wide variety of cells and tissues. Recently, a simplified method of cloning differential display products, separated on agarose gels, was described. Here we report an adaption of this method, using total RNA, to clone differentially expressed genes. The approach is simple and rapid, and requires only small quantities of;total RNA. Utilising this approach, we have cloned three differentially regulated genes from chondrocytes stimulated to hypertrophy in vitro, and confirmed their pattern of expression by Northern blotting. These gene fragments were sequenced and found to correspond to known genes, although only one has previously been isolated from chondrocytes. (C) 1998 Elsevier Science B.V.
引用
收藏
页码:237 / 241
页数:5
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