GUANINE-NUCLEOTIDE EXCHANGE FACTOR FOR EUKARYOTIC INITIATION FACTOR-II - CLONING OF CDNA FOR THE DELTA-SUBUNIT OF RABBIT TRANSLATION INITIATION FACTOR-2B

Peptide sequence data for rabbit eIF-2B delta were obtained and used t o design redundant oligonucleotides for PCR. RNA was isolated from rab bit liver and used to direct the synthesis of total cDNA. A rabbit eIF -2B delta transcript was then amplified by PCR and sequenced. The PCR product was used to isolate a clone from a rabbit liver cDNA library. RACE (rapid amplification of cDNA ends) was used to obtain further 5' sequence. Subsequently, a full length cDNA was obtained from a rabbit reticulocyte library. PCR was used to confirm that the sequence is the same for the liver factor. The sequence obtained shows strong homolog y to that of yeast eIF-2B delta, the GCD2 gene product.