Structure of rodent helix-destabilizing protein revealed by cDNA cloning.

摘要: A cDNA library of newborn rat brain poly(A+) RNA in lambda gt 11 was screened with a synthetic oligonucleotide probe corresponding to five amino acid sequence the N-terminal region calf helix-destabilizing protein, UP1. Six positive phage were isolated after testing 2 X 10(5) recombinants, and each plaque purified. Four these clones second 5 C-terminal UP1; one both probes selected for detailed study. This phage, designated HDP-182, contained 1706-base pair insert an mRNA poly(A) at 3' terminus single open reading frame starting 63 bases from 5' extending 988 bases. The untranslated 718 bases, including AAUAAA signal 21 16-residue poly(U) flanked on side by repeats. Primer extension analysis suggested that HDP-182 full length except about 35 nucleotide residues missing end region, Northern blot revealed relatively abundant species approximately same size as insert. 988-residue predicted 34,215-dalton protein 320 acids. Residues through 196 this are identical 195-residue 124-amino portion is not present purified 124-residue has unusual content it 11% asparagine, 15% serine, 40% glycine consists 16 consecutive oligopeptide Computer-derived secondary structure predictions two distinct domains consisting 1 197 320, respectively.