Molecular Cloning of a Cdna Encoding a Nerve Growth Factor Precursor from the Agkistrodon Halys Pallas

AbstractNGF cDNAs from many sources have been cloned. Here we report a new practical method for cloning a nearly full-length NGF cDNA. Due to the high conservation in the 5’ and 3’ untranslated regions, oligonucleotides complementary to all sequences coding for them were synthesized. Total RNA was extracted from the Agkistrodon halys Pallas venom gland. Accordingly, the first strand of cDNA was synthesized and used as template. The RT-PCR amplified DNA manifested as a single band of predicted size after electrophoresis.Nucleotide sequence analysis revealed an open reading frame of 726 bases encoding a 242 amino acid polypeptide, which includes Agkistrodon halys Pallas prepro-NGF, and whose sequence shows a high similarity to the prepro-NGF of cobra and krait Bungarus multicinctus. Comparison with known sequences shows that the 5’ and 3’ untranslated sequences show slightly higher conservation than the translated region. The prepro part of the precursor is less conserved. The Agkistrodon cDNA possesses the...