| Autor: |
Milojevic, Tetyana, Sonnleitner, Elisabeth, Romeo, Alessandra, Djinović-Carugo, Kristina, Bläsi, Udo |
| Zdroj: |
RNA Biology; June 2013, Vol. 10 Issue: 6 p1066-1069, 4p |
| Abstrakt: |
A His-tag is often added by means of recombinant DNA technology to a heterologous protein of interest, which is then over-produced in Escherchia coliand purified by one-step immobilized metal-affinity chromatography (IMAC). Owing to the presence of 24 histidines at the C-termini of the hexameric E. coliRNA chaperone Hfq, the protein co-purifies with His-tagged proteins of interest. As Hfq can bind to distinct RNA substrates with high affinity, its presence can obscure studies performed with (putative) RNA binding activities purified by IMAC. Here, we present results for a seemingly positive RNA-binding activity, exemplifying that false-positive results can be avoided if the protein of interest is either subjected to further purification step(s) or produced in an E. coli hfq- strain. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
|
