A GENE-EXPRESSION SCREEN

被引:305
作者
WANG, Z
BROWN, DD
机构
关键词
CDNA SUBTRACTION; AMPHIBIAN METAMORPHOSIS; TAIL RESORPTION; GENE EXPRESSION; THYROID HORMONE;
D O I
10.1073/pnas.88.24.11505
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A gene expression screen identifies mRNAs that differ in abundance between two mRNA mixtures by a subtractive hybridization method. The two mRNA populations are converted to double-stranded cDNAs, fragmented, and ligated to linkers for polymerase chain reaction (PCR) amplification. The multiple cDNA fragments isolated from any given gene can be treated as alleles in a genetic screen. Probability analysis of the frequency with which multiple alleles are found provides an estimation of the total number of up- and down-regulated genes. We have applied this method to genes that are differentially expressed in amphibian tadpole tail tissue in the first 24 hr after thyroid hormone treatment, which ultimately induces tail resorption. We estimate that there are about 30 up-regulated genes; 16 have been isolated.
引用
收藏
页码:11505 / 11509
页数:5
相关论文
共 22 条
[1]  
ALT FW, 1978, J BIOL CHEM, V253, P1357
[2]   ISOLATION OF BIOLOGICALLY-ACTIVE RIBONUCLEIC-ACID FROM SOURCES ENRICHED IN RIBONUCLEASE [J].
CHIRGWIN, JM ;
PRZYBYLA, AE ;
MACDONALD, RJ ;
RUTTER, WJ .
BIOCHEMISTRY, 1979, 18 (24) :5294-5299
[3]   INSERTIONAL MUTAGENESIS OF THE DROSOPHILA GENOME WITH SINGLE P-ELEMENTS [J].
COOLEY, L ;
KELLEY, R ;
SPRADLING, A .
SCIENCE, 1988, 239 (4844) :1121-1128
[4]   EXPRESSION OF A SINGLE TRANSFECTED CDNA CONVERTS FIBROBLASTS TO MYOBLASTS [J].
DAVIS, RL ;
WEINTRAUB, H ;
LASSAR, AB .
CELL, 1987, 51 (06) :987-1000
[5]   ISOLATION OF CDNAS OF SCRAPIE-MODULATED RNAS BY SUBTRACTIVE HYBRIDIZATION OF A CDNA LIBRARY [J].
DUGUID, JR ;
ROHWER, RG ;
SEED, B .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (15) :5738-5742
[6]   LIBRARY SUBTRACTION OF INVITRO CDNA LIBRARIES TO IDENTIFY DIFFERENTIALLY EXPRESSED GENES IN SCRAPIE INFECTION [J].
DUGUID, JR ;
DINAUER, MC .
NUCLEIC ACIDS RESEARCH, 1990, 18 (09) :2789-2792
[7]   LOW-RATIO HYBRIDIZATION SUBTRACTION [J].
FARGNOLI, J ;
HOLBROOK, NJ ;
FORNACE, AJ .
ANALYTICAL BIOCHEMISTRY, 1990, 187 (02) :364-373
[8]   ISOLATION OF CDNA CLONES ENCODING T-CELL-SPECIFIC MEMBRANE-ASSOCIATED PROTEINS [J].
HEDRICK, SM ;
COHEN, DI ;
NIELSEN, EA ;
DAVIS, MM .
NATURE, 1984, 308 (5955) :149-153
[9]   IDENTIFICATION OF A SET OF GENES EXPRESSED DURING THE G0/G1 TRANSITION OF CULTURED MOUSE CELLS [J].
LAU, LF ;
NATHANS, D .
EMBO JOURNAL, 1985, 4 (12) :3145-3151
[10]  
LELOUP J, 1977, CR ACAD SCI D NAT, V284, P2261