Open AccessMolecular cloning and the nucleotide sequence of cDNA for neuron-specific enolase messenger RNA of rat brain.
Author(s) -
Kenji Sakimura,
Etsuko Kushiya,
Masuo Obinata,
Shoji Okada,
Yasuo Takahashi
Publication year - 1985
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.82.21.7453
Subject(s) - complementary dna , microbiology and biotechnology , biology , messenger rna , coding region , nucleic acid sequence , southern blot , cdna library , enolase , northern blot , genomic dna , restriction enzyme , rna , dot blot , dna , genetics , gene , immunohistochemistry , immunology
The cDNAs to mRNA for rat gamma gamma enolase (neuron-specific enolase; NSE; EC 4.2.1.11) were isolated from a cDNA library by using differential colony hybridization and a hybrid-selected translation assay. By overlapping of the nucleotide sequences of several cDNA inserts, it was found that they spanned 2232 base pairs (bp) which included 1299 bp of the complete coding region, 68 bp of the 5' noncoding region, and 848 bp of the 3' noncoding region, including a polyadenylylation signal. In addition, the poly(A) tail was also found. The amino acid sequence deduced from the nucleotide sequence was composed of 433 amino acids. Southern blot analysis with a cDNA insert detected one hybridizing fragment in rat genomic DNA digested with several different restriction enzymes. Dot-blot and transfer hybridization analyses of poly(A)+ RNA from developing rat brains showed an increase of NSE mRNA 10-30 days after birth.