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cDNA cloning and expression of stress‐inducible rat hsp70 normal and injured rat brain
Author(s) -
Longo F. M.,
Wang S.,
Narasimhan P.,
Zhang J. S.,
Chen J.,
Massa S. M.,
Sharp F. R.
Publication year - 1993
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/jnr.490360310
Subject(s) - complementary dna , biology , microbiology and biotechnology , hsp70 , messenger rna , northern blot , open reading frame , reverse transcriptase , gene expression , blot , rapid amplification of cdna ends , reverse transcription polymerase chain reaction , cloning (programming) , heat shock protein , gene , rna , molecular cloning , peptide sequence , biochemistry , computer science , programming language
A reverse transcriptase‐polymerase chain reaction(RT‐PCR) product obtained from ischemic rat brain RNA was used to screen a rat ischemic forebrain cDNA library for a cDNA clone containing the entire open reading frame for the inducible hsp70. The coding sequence for the rat hsp70 cDNA demonstrated significant similarities with the human hsp70 of Hunt and Morimoto ( Proc Natl Acad Sci 82:6455–6459, 1985) and the mouse hsp70 of Hunt and calderwood ( Gene 87:199–204, 1990). The rat inducible hsp70 and constitutive hsc73 sequences are distinct. There was a low level of hsp70 mRNA expression in normal rat brain as is found in other tissues. hsp70 mRNA was markedly induced in rat brain 8 hours following global ischomia and kainic acid‐induced sizures. Northern blots showed a ∼2.9kb hsp70 mRNA band from control, kainic acid, and ischemic brains. RTPCR confirmed the presence of hsp70 mRNA in normal rat brain. Since there are at least five human and six mouse inducible hsp70 genes known, many other rat hsp70 genes probably exist that could function in different cells or organelles or be induced under different circumstances. © 1993 Wiley‐Liss, Inc.