Open AccessLibrary subtraction ofin vitrocDNA libraries to identify differentially expressed genes in scrapie infection
Author(s) -
John R. Duguid,
Mary C. Dinauer
Publication year - 1990
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/18.9.2789
Subject(s) - biology , cdna library , complementary dna , primer (cosmetics) , microbiology and biotechnology , in vitro , polymerase chain reaction , subtraction , scrapie , genomic library , gene , virology , genetics , base sequence , chemistry , organic chemistry , prion protein , medicine , arithmetic , mathematics , disease , pathology
We have developed a system where double-stranded cDNA can be amplified using a synthetic oligonucleotide primer and the polymerase chain reaction, generating cDNA libraries in vitro. Using a library subtraction strategy (1), scrapie and control brain in vitro cDNA libraries were used to identify sequences whose expression is modulated in scrapie infection. One of these sequences represents beta-2 microglobulin, while the other two have not been previously described. The use of in vitro libraries offers increased speed and efficiency of construction, and their subtraction is more efficient and powerful, compared with the previous system (1).