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Isolation and characterization of unique sequence DNA clones corresponding to rare transcripts from rat brain
Author(s) -
Brilliant M.H.,
Sueoka N.,
Chikaraishi D.M.
Publication year - 1983
Publication title -
international journal of developmental neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.761
H-Index - 88
eISSN - 1873-474X
pISSN - 0736-5748
DOI - 10.1016/0736-5748(83)90319-2
Subject(s) - dept , citation , library science , special collections , presentation (obstetrics) , isolation (microbiology) , computer science , biology , genetics , medicine , bioinformatics , radiology
A genomic l i b ra r y of s ingle copy rat DNA was prepared in phage Ml3. Total rat DNA was digested with the res t r i c t i on endonuclease Mbol. Unique Sequence DNA was isolated and l igated in to the Bam HI s i te of the rep l i ca t i ve form of the bacteriophage Ml3 mp7. Single stranded DNA from ind iv idua l recombinant Ml3 mp7 clones was digested with Sau 3A, which p re fe ren t i a l l y removes the rat DNA insert , label led with 32p by polynucleotide kinase, and hybridized to adult brain to ta l RNA. Clones containing sequences homologous to brain t ranscr ip ts were determined by a var ia t ion of the t ranscr ip t ion mapping technique. About 20% of these s ingle stranded clones were homologous to adult rat brain RNA, which agrees with our ea r l i e r RNA complexity data (Chikaraishi et a l , Cell 13, 1978). Also as expected, these clones correspond to rare brain t ranscr ip ts (O.05-1~opy per c e l l ) . These sequences were fu r ther characterized in terms of t ranscr ip t ion in heterologous tissues and in new born brain. At least two of the clones are spec i f i ca l l y transcribed in brain. The t ranscr ip t ion patterns of these cloned DNA fragments may reveal developmental patterns of gene expression.