Cloning and partial sequencing of a new rat brain specific cDNA.
In order to find brain specific transcripts in a pUC13 cDNA library prepared from rat brain cytoplasmic poly(A)+ RNAs, the following steps were observed: 1. Randomly chosen cDNA clones from the brain library were screened by radiolabelled single stranded cDNAs (sscDNAs) prepared from liver, spleen, kidney and intestine mRNAs. 2. The brain clones containing genetic information shared with the peripheral organs were discarded. 3. After hybridization of the remaining clones with radiolabelled sscDNAs prepared from rat brain poly(A)+ RNAs, 210 possibly brain specific clones were selected. 4. Plasmids containing cDNA of each of these clones were purified and after estimation of their cDNA sizes, clones containing inserts of at least 500 base pairs (bp) were used as radiolabelled probes against rat brain, liver, spleen, kidney and intestine poly(A)+ RNAs spotted onto nitrocellulose filters (dot blot analysis). 5. By using this screening procedure, several brain specific clones were obtained. One of them (clone III 25) is presently subjected to sequencing in M13 phage. 6. A computer analysis of the partial sequence so far obtained gives no significant homology to the 15.000 genes contained in Genebank and EMBL data bank. It seems therefore that we have isolated a new brain specific cDNA coding for a not yet described peptide.
Marechal
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Laboratoire de Pharmacologie, Université de Liège, Belgique.
Delapierre
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Dresse
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D000818:Animals; D001483:Base Sequence; D001923:Brain Chemistry; D003001:Cloning, Molecular; D004247:DNA; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D009928:Organ Specificity; D012333:RNA, Messenger; D051381:Rats; D012689:Sequence Homology, Nucleic Acid
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; D004247:DNA