Title

Cloning and over expression of non-coding RNA rprA in E. coli and its resistance to Kanamycin
without osmotic shock

Authors

Azita Sahni¹, Mohammadreza Hajjari^2*, Jamshid Raheb^3*, Ali Mohammad Foroughmand²,
Morteza Asgari¹

Affiliation

¹Nour Danesh Institute of Higher Education, Department of Biology, Isfahan, Iran;

²Department of Genetics, Faculty of Science, Shahid Chamran University of Ahvaz, Ahvaz, Iran;

³National Institute of Genetic Engineering and Biotechnology, Tehran, Iran;
 

Email

M-hajari@scu.ac.ir;

jamshid@nigeb.ac.ir;

Article Type

Hypothesis

Date

November 8, 2016, Revised December 2, 2016; Accepted December 2, 2016; Published January 31, 2017

Recent reports have indicated that small RNAs have key roles in the response of the E.coli to stress and also in the regulating of virulence factors. It seems that some small non-coding RNAs are involved in multidrug resistance. Previous studies have indicated that rprA can increase the tolerance to Kanamycin in RcsB-deficient Escherichia coli K-12 following osmotic shock. The current study aims to clone and over-express the non-coding RNA rprA in E.coli and investigate its effect on the bacterial resistance to Kanamycin without any osmotic shock. For this purpose, rprA gene was amplified by the PCR and then cloned into the PET-28a (+) vector. The recombinant plasmid was transformed into wild type E.coli BL21 (DE3). The over expression was induced by IPTG and confirmed by qRT-PCR. The resistance to the kanamycin was then measured in different times by spectrophotometry. The statistical analysis showed that the rprA can increase the resistance to Kanamycin in Ecoli K12. The interaction between rprA and rpoS was reviewed and analyzed by in silico methods. The results showed that the bacteria with over-expressed rprA were more resistance to Kanamycin. The present study is an important step to prove the role of non-coding RNA rprA in bacterial resistance. The data can be the basis for future works and can also help to develop and deliver next-generation antibiotics.

Keywords

Escherichia coli, non-coding RNA, rprA, Kanamycin

Citation

Sahni et al. Bioinformation 13(1): 21-24 (2017)

Edited by

P Kangueane

ISSN

0973-2063

Publisher

Biomedical Informatics

License

This is an Open Access article which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. This is distributed under the terms of the Creative Commons Attribution License.