High-level expression of the bovine growth hormone gene in heterologous mammalian cells.
The gene coding for bovine growth hormone (bGH) was isolated from a lambda-phage library constructed using bovine pituitary DNA partially digested with MboI. Expression of this gene transfected into mouse and monkey cells was studied. CV-1 monkey cells transfected with simian virus 40 (SV40) vectors containing the intact bGH gene, including the putative promoter region, did not express bGH. However, replacement of the bGH promoter with the mouse metallothionein-I (MT) promoter resulted in high-level synthesis and secretion of bGH. These results show that the bGH promoter functions poorly in CV-1 cells but CV-1 cells process and translate the bGH mRNA accurately. The MT-bGH chimeric gene was used to establish permanent bGH-secreting mouse C127 cell lines using the 69% transforming fragment of bovine papilloma virus (BPV) as the vector. One such cell line produced high levels of bGH and secreted it into the medium efficiently. Secreted bGH is processed accurately and is bioactive as judged by its ability to bind to rabbit liver membrane preparations.
D000818:Animals; D001681:Biological Assay; D002417:Cattle; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D002874:Chromosome Mapping; D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D013006:Growth Hormone / Q000235:genetics*; D008668:Metallothionein / Q000235:genetics; D051379:Mice; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational; D012323:RNA Processing, Post-Transcriptional
D004274:DNA, Recombinant; D013006:Growth Hormone; D008668:Metallothionein; D004262:DNA Restriction Enzymes