RECOMBINANT PLASMID OF ESCHERICHIA COLI FOR AMPLIFICATION OF HUMAN GROWTH HORMONE COMPLEMENTARY STRAND DNA

摘要

PURPOSE:To enable the mass-production of human growth hormone gene, by forming an established human pregrowth hormonegene-amplifying plasmid originated from human pituitary tissue and having a specific deoxynucleotide linkage. CONSTITUTION:Human putuitary tissue extracted from human is disintegrated in a guanidine thiocyanate solution, and centrifuged to collect the RNA fraction, which is passed through an oligo-dT column to concentrate the messenger RNA having poly A terminal. The double chain containing 5'-terminal poly dC linkage which is not at the side of complementary DNA is removed by the treatment with a restriction enzyme Hind III, and the residue is added with a part of the SV40 gene containing about 260 base pairs and having about 15 poly dG linkages and a Hind III adhesive end at the 5'-terminal. The product is made to react with RNase H, DNA-polymerase and DNA-ligase to obtain the objective plasmid having complementary DNA of various messenger RNAs.