Facile cloning and sequencing of S-crystallin genes from octopus lenses based on polymerase chain reaction

Abstract

S-crystallin is a major lens protein present in the octopus and squid of Cephalopods. To facilitate the cloning of the protein, cDNA was constructed from the poly(A)+RNA of octopus lenses, and amplification by polymerase chain reaction (PCR) was carried out with two primers designed according to the 5'- and 3'-coding regions of S-crystallin gene. Sequencing two of 15 positive clones obtained shows 37-44% similarity in nucleotide and 23-30% similarity in amino acid sequences as compared with mammalian glutathione S-transferases (GST), revealing that S-crystallins exist as a multigene family and probably derived from GST by gene duplication and subsequent mutational base replacements.