RECOMBINANT PLASMID OF ESCHERICHIA COLI FOR AMPLIFICATION OF HUMAN PROLACTIN COMPLEMENTARY STRAND DNA

摘要

PURPOSE:To enable the mass-production of human prolactin gene, by forming an established human preprolactingene-amplifying plasmid originated from human pituitary tissue and having a specific deoxynucleotide linkage. CONSTITUTION:An auxiliary DNA of poly A RNA is synthesized by a reverse transcriptase, and about 15 poly dC linkages are attached to the 5'-terminal with a deoxinucleotide terminal translocase. The double chain containing the 5'-terminal poly dC linkage which is not at the side of complementary DNA is removed by treating with a restriction enzyme Hind III, and the residue is added with a part of the SV40 gene containing about 280 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 cyclized messenger RNAs.