Roles of neurotrophic factors derived from astrocyte in neuronal development

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(1) Cloning of human GMF-gamma cDNAWe isolated a human GMF-gamma cDNA from human fetal brain cDNA library. One of the clones contained a 573 nucleotide long insert and an open reading frame of 426 bp encoding a deduced protein of 142 amino acids.(2) Expression of human GMF-beta and GMF-gammaThe cDNA of human GMF-beta and GMF-gamma were inserted to pAED4 expression vector and introduced to E. coli BL21(DE3). The expressed proteins were purified and subjected to antibody production.(3) Production of anti-human GMF-beta and GMF-gamma antibodiesRecombinant human GMF-beta and GMF-gamma were used as immunogens for the production of polyclonal antibodies. The each anti-serum recognized human GMF-beta and GMF-gamma respectively, but either of them did not react to rat GMF-beta or GMF-gamma.(4) Cloning of rat GMF-gamma cDNAWe isolated a rat GMF-gamma cDNA from rat pulmonary artery cDNA library. One of the clones contained a 553 nucleotide long insert and an open reading frame of 426 bp encoding a deduced protein of 142 amino acids.(5) Expression of rat GMF-beta and GMF-gammaThe cDNA of rat GMF-beta and GMF-gamma were inserted to pAED4 expression vector and introduced to E. coli BL21(DE3). The expressed proteins were purified and subjected to antibody production.(6) Production of anti-rat GMF-beta and GMF-gamma antibodiesAnti-rat GMF-beta and GMF-gamma serum were produced by the same way of human GMF-beta and GMF-gamma. The anti-sera recognized rat GMF-beta and GMF-gamma respectively.