Molecular beacon aptamers for direct and universal quantitation of recombinant proteins from cell lysates.
Western blot, enzyme linked immunosorbent assay (ELISA), and fluorescent fusion proteins are currently the most common methods for detecting recombinant proteins. However, the former two are cumbersome and time-consuming, and the latter method may interfere with the trafficking and function of the fused recombinant proteins. We report here a rapid, inexpensive, and simple approach to detect and quantify recombinant proteins using an anti-His-tag molecular beacon aptamer (HMBA). We demonstrated the technique by detection and quantitation of expressed recombinant proteins directly from E. coli cell lysate. The amount of expressed P78-His was determined to be 1.49 μg from the 20 μg cell lysate proteins. To the best of our knowledge, this is the first example directly measuring the concentration and expression yield of recombinant proteins from cell lysate, and the entire procedure required only 5 min.
Molecular Science and Biomedicine Laboratory, College of Biology, Hunan University, Changsha, 410082, P. R. China.
Xiaohong
X
Tan
Weijun
W
Chen
Shun
S
Lu
Zhi
Z
Zhu
Tao
T
Chen
Guizhi
G
Zhu
Mingxu
M
You
Weihong
W
Tan
D052157:Aptamers, Nucleotide / Q000737:chemistry*; D004926:Escherichia coli / Q000166:cytology* / Q000235:genetics; D018398:Homeodomain Proteins / Q000032:analysis* / Q000235:genetics; D006801:Humans; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics; D015534:Trans-Activators / Q000032:analysis* / Q000235:genetics
D052157:Aptamers, Nucleotide; D018398:Homeodomain Proteins; D011994:Recombinant Proteins; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein