公开(公告)号：EP1309604A4

公开(公告)日：2006-10-11

申请号：EP01964014

申请日：2001-08-15

申请人： PHAGE BIOTECHNOLOGY CORP

发明人： KORDYUM VITALIY A ,  CHERNYKH SVITLANA I ,  SLAVCHENKO IRYNA YU ,  VOZIANOV OLEKSANDR F

IPC分类号： C12N15/09 ,  C07K14/50 ,  C12N9/52 ,  C12N15/63 ,  C12N15/70 ,  C12P21/02 ,  C07H21/04 ,  C12N15/73 ,  C12P21/00

CPC分类号： C12P21/02 ,  C07K14/50 ,  C12N15/63 ,  C12N15/70

摘要： This invention relates to a method for enhancing the production of biologically active proteins and peptides in bacterial cells by infecting bacterial cells of the producer strain, which contain a plasmid with one or more targeted genes, with bacteriophage μ with or without the targeted gene(s). The targeted genes encoding the biologically active proteins are under the control of a T7 polymerase promoter and the bacteria also are capable of expressing the gene for T7 RNA polymerase. The phage increases synthesis of the targeted protein and induces lysis of the producer strain cells. Super-production is achieved by the combination of the high level of expression achieved from the T7 polymerase promoter and by cultivating the producer strain cells under culture conditions that delay lytic development of the phage. The biologically active proteins and peptides subsequently accumulate in a soluble form in the culture medium as the cells of the producer strain are lysed by the phage.

公开(公告)号：EP1309716A4

公开(公告)日：2005-08-03

申请号：EP01967977

申请日：2001-08-15

申请人： PHAGE BIOTECHNOLOGY CORP

发明人： STEGMANN THOMAS J ,  KORDYUM VITALIY A ,  CHERNYKH SVITLANA I ,  SLAVCHENKO IRYNA YU ,  VOZIANOV OLEKSANDR F

IPC分类号： C12N15/09 ,  A61K38/00 ,  A61P9/00 ,  C07K14/50 ,  C12N1/21 ,  C12N7/01 ,  C12N15/70 ,  C12N15/73 ,  C12P21/02 ,  C12P21/00 ,  C07H21/04 ,  C12N7/00

CPC分类号： C07K14/501 ,  C12N15/70

摘要： The gene of human acidic fibroblast growth factor 155 (haFGF 155) has been obtained by chemical synthesis. The nucleotide sequence of haFGF 155 gene has been deduced on the basis of haFGF 155 amino acid sequence as described in the literature. The amino acid sequence, of the synthesized haFGF 155 does not differ from those described in the literature. The nucleotide sequence of haFGF gene differs from those described previously. For chemical synthesis of haFGF 155 gene, codons were used which are the ones most often used by E. coli in highly expressed E. coli proteins. A plasmid with haFGF 155 (phaFGF 155) gene was obtained and was used to transform E. coli. Production of haFGF 154 protein was achieved by cultivation of the producer strain under conditions which slow down the lytic development of lambda phage. The haFGF 154 protein accumulated in culture medium in a soluble condition as a result of the producer strain cells lysis by the lambda phage. The haFGF 154 protein constituted 20% of the soluble protein accumulated in the culture medium and its biological activity was demonstrated by its ability to generate new vessels (angiogenesis). The initiator methionine residue at position 1 of the FGF 155 protein was completely removed during protein synthesis resulting in an FGF 154 amino acid product. The use of the phage-dependent method to produce other forms of the haFGF protein is also disclosed.