公开(公告)号：WO2014066404A3

公开(公告)日：2014-08-07

申请号：PCT/US2013066204

申请日：2013-10-22

Applicant: LIFE TECHNOLOGIES CORP

Inventor： ROGERS JEFFREY ,  FLETCHER JULIA ,  LOWITZ KEVIN

IPC: C12N15/10 ,  C12N15/62

CPC classification number: C12N15/67 ,  C07K2/00 ,  C07K2319/40 ,  C07K2319/60 ,  C12N9/2471 ,  C12N15/1086 ,  C12N15/63 ,  C12N15/70 ,  C12P21/00

Abstract: 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 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.

Abstract translation: 本公开描述了新的重组基因表达系统及其使用方法。 重组基因表达系统适合于使用户能够通过允许一小部分（例如，少于10％）的靶蛋白被表达为与易于检测的报道分子的融合物来实时监测蛋白质的成功表达 。 通过提供具有核苷酸序列N4R6NX的随机核糖体结合位点，其中N是A，T，G或C，其中R是A，所述重组基因表达系统还允许用户选择表现高，中或低表达的克隆 或G，并且其中x是克隆基因上游6至11的整数。 通过选择展现出期望的报道基因表达相对水平的克隆，用户可以调整所需蛋白质的表达水平。