PURPOSE: To obtain a hybrid plasmid capable of producing natural-type hGH by binding a cloning vector of Bacillus subtilis to a gene coding hGH fused on the 5' end to a sequence coding serine protease's secretion signal.
CONSTITUTION: First, a cloning vector of Bacillus subtilis, e.g. ATCC 67320p SM214, is cut off by restriction enzymes Ecol I and Hind III to separate EcoR I/Hind III fragment having 6500 base pairs. Secondly, plasmid pSM209 is cut off by restriction enzymes FnuD II and Hind III to separate FnuD II/Hind III fragment with 530 base pairs coding hGH's 17-191 amino acid sequence. Subsequently, an oligouncleotide coding a signal sequence responding to serine protease secretion and hGH's 1-16 amino acid sequence is synthesized, and the above respective DNA fragments are mutually bound via T4 DNA ligase, thus obtaining the objective hybrid plasmid capable of inducing the expression and secretion of natural-type hGH.
ERIZABETSUTA FURANKI
FUEDERIIKO MAIZAANO
SHIRUBIA ASUTORUA TESUTOORI
| JPS62163691A | 1987-07-20 | |||
| JPS6070075A | 1985-04-20 |