A cDNA library was prepared from mRNA extracted from bovine pituitaries,β-subunitDNA of bFSH was amplified from cDNA using Polymerase Chain Reaction(PCR).The sequence of aminoacid encoded by cDNA was the same as that...A cDNA library was prepared from mRNA extracted from bovine pituitaries,β-subunitDNA of bFSH was amplified from cDNA using Polymerase Chain Reaction(PCR).The sequence of aminoacid encoded by cDNA was the same as that of natural bFSH by sequencing.Signal peptide DNAfragment of bFSH was deleted by PCR mediated mutagenesis,to which PGEX-2T was ligated,yielding a high expression fusion protein of 40 kD.A recombinant bFSH fusion protein was extracted usingthrombin,yielding a pure bFSH of 14kD,showing positive reaction using Western blot analysis,and someactivation by biological tests with the mouse and the rabbit.展开更多
文摘A cDNA library was prepared from mRNA extracted from bovine pituitaries,β-subunitDNA of bFSH was amplified from cDNA using Polymerase Chain Reaction(PCR).The sequence of aminoacid encoded by cDNA was the same as that of natural bFSH by sequencing.Signal peptide DNAfragment of bFSH was deleted by PCR mediated mutagenesis,to which PGEX-2T was ligated,yielding a high expression fusion protein of 40 kD.A recombinant bFSH fusion protein was extracted usingthrombin,yielding a pure bFSH of 14kD,showing positive reaction using Western blot analysis,and someactivation by biological tests with the mouse and the rabbit.
