System and Method for Visualization of Optimized Protein Expression

摘要

The present disclosure describes a novel recombinant gene expression system and a method of its use. The recombinant gene expression system is adapted to enable a user to monitor the successful expression of a protein in real time by allowing a small portion (e.g., less than 10%) of the target protein to be expressed as a fusion with a readily detectable reporter. The recombinant gene expression system further allows a user to select a clone that exhibits high, medium or low expression by providing a random ribosomal binding site having the nucleotide sequence N4R6NX, where N is A, T, G, or C, where R is A or G, and where x is an integer from 6 to 11 (SEQ ID NO: 1) upstream of the cloned gene. By selecting a clone exhibiting a desired relative level of reporter gene expression, a user can tailor the expression level of the desired protein.

主权项

1. An expression vector comprising:
a promoter capable of driving transcription in a host cell; a random ribosomal binding site (RRBS) functionally linked to and positioned downstream from the promoter; a stop codon context element comprising a stop codon and at least onel 3-nucleotide codon in frame with the stop codon; and at least a portion of a gene encoding a reporter protein downstream of and in-frame with the stop codon context element.