| 摘要 |
PURPOSE:To obtain a precursor of human macrophage chemotactic factor in high purity and large quantities by preparing a now gene from mRNA derived from a human peripheral blood lymphocyte, human T lymphocyte hybridoma and/or human T lymphocyte based established cell. CONSTITUTION:A normal human peripheral blood lymphocyte, human T human lymphocyte established cell and/or cloning strain is cultured and a cell capable of synthesizing a protein having an amino sequence having a human macrophage chemotactic factor(MCF) or partial structure thereof is cultured therefrom and a fraction containing mRNA is separated from total RNA fraction obtained by grinding the resultant cell and extracting from the ground cell. Then cDNA prepared using the above-mentioned mRNA as template and successively using a reverse transcription enzyme, reverse transcriptase and ribonuclease and DNA synthetic enzyme is integrated into a vector and the resultant recombinant is introduced into a host to provide cDNA library. The cDNA coding a human MCF precursor protein is cloned from the above-mentioned cDNA library by a plaque hybridization method to provide the cloned DNA capable of coding a human MCF precursor protein and having a base sequence expressed by the formula. |
