Expression of soluble heterologous proteins in bacteria utilizing a thioredoxin/protein expression vector
View Patent ↗A method for producing a soluble, properly folded, biologically active protein in bacteria in which the heterologous protein is coexpressed with a thioredoxin protein.
1. A method for expressing a soluble heterologous protein in bacteria comprising: transforming a bacterium with a vector wherein the vector contains a nucleic acid sequence which expresses thioredoxin and a nucleic acid sequence which expresses a heterologous protein; and culturing the bacterium under conditions wherein the thioredoxin and the heterologous protein are expressed as separate non-fused proteins and the heterologous protein is expressed in a soluble, biologically active form.
2. The method of claim 1 wherein the vector is a plasmid.
3. The method of claim 1 wherein the nucleic acid sequence which expresses thioredoxin and the nucleic acid sequence which expresses the heterologous protein are operationally linked to a common promoter.
4. The method of claim 3 wherein the promoter is a lac promoter.
5. A vector which contains a nucleic acid sequence which encodes a thioredoxin protein and a nucleic acid sequence which encodes a heterologous protein wherein the vector expresses the thioredoxin protein and the heterologous protein as separate, nonfused proteins, and wherein the heterologous protein is expressed in a soluble, biologically active form.
6. The vector of claim 5 wherein the vector is a plasmid.
7. The vector of claim 5 wherein the nucleic acid sequence which encodes the thioredoxin protein and the nucleic acid sequence which encodes the heterologous protein are operationally linked to a common promoter.
8. The vector of claim 7 wherein the promoter is a lac promoter.
9. A bacterium transformed with an expression vector containing a nucleic acid encoding a thioredoxin protein and a nucleic acid encoding a heterologous protein wherein the thioredoxin protein and the heterologous protein are expressed as separate proteins and wherein the heterologous protein is expressed in a soluble, biologically active form.
10. The bacterium of claim 9 wherein the bacterium is Escherichia coli.
11. The bacterium of claim 9 wherein the vector is a plasmid.
12. The bacterium of claim 9 wherein the nucleic acid which encodes the thioredoxin and the nucleic acid which encodes the heterologous protein are operationally linked to a common promoter.
13. The bacterium of claim 12 wherein the promoter is a lac promoter.