Isolation of cDNAs of scrapie-modulated RNAs by subtractive hybridization of a cDNA library.
We have developed a subtractive cloning procedure based on the hybridization of single-stranded cDNA libraries constructed in pi H3M, a vector containing the phage M13 origin of replication. We have used this strategy to isolate three transcripts whose abundance is increased in scrapie-infected brain. DNA sequence analysis showed that they represent glial fibrillary acidic protein, metallothionein II, and the B chain of alpha-crystallin; the latter two may represent a response to stress.
Duguid
J R
JR
GRECC, Edith Nourse Rogers Memorial Veterans Administration Hospital, Bedford, MA 01730.
Rohwer
R G
RG
Seed
B
B
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry; D003001:Cloning, Molecular; D006224:Cricetinae; D003459:Crystallins / Q000235:genetics; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D005260:Female; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics; D008647:Mesocricetus; D008668:Metallothionein / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012313:RNA / Q000235:genetics*; D012608:Scrapie / Q000235:genetics*; D014158:Transcription, Genetic
D003459:Crystallins; D005904:Glial Fibrillary Acidic Protein; D012313:RNA; D004247:DNA; D008668:Metallothionein