United States Patent
US Patent 4237224: Process for producing biologically functional…
US 4237224 · granted 1980-12-02
Abstract
Method and compositions are provided for replication and expression of exogenous genes in microorganisms. Plasmids or virus DNA are cleaved to provide linear DNA having ligatable termini to which is inserted a gene having complementary termini, to provide a biologically functional replicon with a desired phenotypical property. The replicon is inserted into a microorganism cell by transformation. Isolation of the transformants provides cells for replication and expression of the DNA molecules present in the modified plasmid. The method provides a convenient and efficient way to introduce genetic capability into microorganisms for the production of nucleic acids and proteins, such as medically or commercially useful enzymes, which may have direct usefulness, or may find expression in the production of drugs, such as hormones, antibiotics, or the like, fixation of nitrogen, fermentation, utilization of specific feedstocks, or the like.
| Patent Number | 4237224 |
|---|---|
| Title | Process for producing biologically functional molecular chimeras |
| Filed | 1979-01-04 |
| Granted | 1980-12-02 |
| Inventor(s) | Boyer; Herbert W., Cohen; Stanley N. |
| Assignee | Board of Trustees of the Leland Stanford Jr. University |
| CPC Classification | C12N 15/00 |
| Number of Claims | 15 |
Abstract
Method and compositions are provided for replication and expression of exogenous genes in microorganisms. Plasmids or virus DNA are cleaved to provide linear DNA having ligatable termini to which is inserted a gene having complementary termini, to provide a biologically functional replicon with a desired phenotypical property. The replicon is inserted into a microorganism cell by transformation. Isolation of the transformants provides cells for replication and expression of the DNA molecules present in the modified plasmid. The method provides a convenient and efficient way to introduce genetic capability into microorganisms for the production of nucleic acids and proteins, such as medically or commercially useful enzymes, which may have direct usefulness, or may find expression in the production of drugs, such as hormones, antibiotics, or the like, fixation of nitrogen, fermentation, utilization of specific feedstocks, or the like.
Claim 1
A method for replicating a biologically functional DNA, which comprises: transforming under transforming conditions compatible unicellular organisms with biologically functional DNA to form transformants; said biologically functional DNA prepared in vitro by the method of: (a) cleaving a viral or circular plasmid DNA compatible with said unicellular organism to provide a first linear segment having an intact replicon and termini of a predetermined character; (b) combining said first linear segment with a second linear DNA segment, having at least one intact gene and foreign to said unicellular organism and having termini ligatable to said termini of said first linear segment, wherein at least one ofsaid first and second linear DNA segments has a gene for a phenotypical trait, under joining conditions where the termini of said first and second segments join to provide a functional DNA capable of replication and transcription in said unicellularorganism; growing said unicellular organisms under appropriate nutrient conditions; and isolating said transformants from parent unicellular organisms by means of said phenotypical trait imparted by said biologically functional DNA.
Claims
15 totalA method for replicating a biologically functional DNA, which comprises: transforming under transforming conditions compatible unicellular organisms with biologically functional DNA to form transformants; said biologically functional DNA prepared in vitro by the method of: (a) cleaving a viral or circular plasmid DNA compatible with said unicellular organism to provide a first linear segment having an intact replicon and termini of a predetermined character; (b) combining said first linear segment with a second linear DNA segment, having at least one intact gene and foreign to said unicellular organism and having termini ligatable to said termini of said first linear segment, wherein at least one ofsaid first and second linear DNA segments has a gene for a phenotypical trait, under joining conditions where the termini of said first and second segments join to provide a functional DNA capable of replication and transcription in said unicellularorganism; growing said unicellular organisms under appropriate nutrient conditions; and isolating said transformants from parent unicellular organisms by means of said phenotypical trait imparted by said biologically functional DNA.