公开(公告)号：US20150267209A1

公开(公告)日：2015-09-24

申请号：US14437317

申请日：2013-10-22

Applicant: Life Technologies Corporation

Inventor： Jeffrey Rogers ,  Julia Fletcher ,  Kevin Lowitz

IPC: C12N15/67 ,  C12N9/38 ,  C07K2/00 ,  C12P21/00 ,  C12N15/70

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 (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.

Abstract translation: 本公开描述了一种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于使用户能够通过使目标蛋白质的一小部分（例如，小于10％）表达为与容易检测的报道分子的融合物来实时监测蛋白质的成功表达 。 重组基因表达系统还允许用户通过提供具有核苷酸序列N4R6NX的随机核糖体结合位点来选择表现出高，中或低表达的克隆，其中N是A，T，G或C，其中R是A 或G，并且其中x是克隆基因上游的6至11（SEQ ID NO：1）的整数。 通过选择表现出所需相对水平的报告基因表达的克隆，用户可以调整所需蛋白质的表达水平。