Determination of Globin Beta-chains and Comparison of Genetic Type of SMMC/B Mice of Three Generations by Peptide Map Analysis / 第二军医大学学报

ABSTRACT

The separation and purification of beta chains of globm from the inbreeding SMMC/B mice were performed by the dissociation of chains in urea, CMhcellulose column chromatography for the separation of alpha and beta chains, and Sephadex G25 column chromatography for removing salts. The pure beta-chain was digested by trypsin. Digested fragments were resolved by cellulose membrane two-way electrophoresis, thus obtaining the map of spots of beta-chain fragments for B33, B34, Bp(), Bp and Bf. The spots,BTp3, which have the same positions ,were separated and micro-sequences were assayed by DABITC/PITC double coupling technique. According to the genetic code, we might deduce the nucleotide sequence of their mRNA from the known amino acid sequence of BTp3, a piece of beta-chain. By analyzing the altered genetic loci of beta-chain of globin, we might determine the genetic purity of the strain and distinguish objectively the genetic types between species and subspecies.