Site-Directed Mutagenesis for In Vitro and In Vivo Experiments Exemplified with RNA Interactions in Escherichia Coli

Journal of Visualized Experiments

Volumn 2019, Issue 144, 2019, Pages

  Andreassen, Patrick Rosendahl ^a   Pettersen, Jens Sivkær ^a   Jørgensen, Mikkel ^a  

^a UNIVERSITY OF SOUTHERN DENMARK   (Denmark)

Author keywords

Biochemistry; dual plasmid; EMSA; Issue 144; Mutagenesis; Mutagenic analysis; oligonucleotide directed mutagenesis; RNA protein interactions; RNA RNA interactions; site directed; western blot

Indexed keywords

RNA;

ESCHERICHIA COLI; GENETICS; METABOLISM; PROCEDURES; SITE DIRECTED MUTAGENESIS;

ESCHERICHIA COLI; MUTAGENESIS, SITE-DIRECTED; RNA;

EID: 85062092809     PISSN: 1940087X     EISSN: None     Source Type: Journal    
DOI: 10.3791/58996     Document Type: Article

References (14)

1. Bouvier, M. et al. Small RNA binding to 5' mRNA coding region inhibits translational initiation. Molecular Cell. 32 (6), 827-837 (2008).
2. Jorgensen, M. G. et al. Small regulatory RNAs control the multi-cellular adhesive lifestyle of Escherichia coli. Molecular Microbioly. 84 (1), 36-50 (2012).
3. Antal, M. et al. A small bacterial RNA regulates a putative ABC transporter. The Journal of Biological Chemistry. 280 (9), 7901-7908 (2005).
4. Andreassen, P. R. et al. sRNA-dependent control of curli biosynthesis in Escherichia coli: McaS directs endonucleolytic cleavage of csgD mRNA. Nucleic Acids Research. (2018).
5. Thomason, M. K. et al. A small RNA that regulates motility and biofilm formation in response to changes in nutrient availability in Escherichia coli. Molecular Microbioly. 84 (1), 36-50 (2012).
6. Lorenz, T. C. Polymerase chain reaction: basic protocol plus troubleshooting and optimization strategies. Journal of Visualized Experiments. (63), e3998 (2012).
7. JoVE Science Education Database. Basic Methods in Cellular and Molecular Biology. Molecular Cloning. Journal of Visualized Experiments. Cambridge, MA (2018).
8. JoVE Science Education Database. Basic Methods in Cellular and Molecular Biology. The Western Blot. Journal of Visualized Experiments. Cambridge, MA (2018).
9. JoVE Science Education Database. Biochemistry. Electrophoretic Mobility Shift Assay (EMSA). Journal of Visualized Experiments. Cambridge, MA (2018).
10. Kunkel, T. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proceedings of the National Academy of Sciences of the United States of America. 82 (2), 488-492 (1985).
11. Laible, M., Boonrod, K. Homemade site directed mutagenesis of whole plasmids. Journal of Visualized Experiments. (27) (2009).
12. Wells, J. A., Vasser, M., Powers, D. B. Cassette mutagenesis: an efficient method for generation of multiple mutations at defined sites. Gene. 34 (2-3), 315-323 (1985).
13. Cong, L. et al. Multiplex genome engineering using CRISPR/Cas systems. Science. 339 (6121), 819-823 (2013).
14. Mali, P. et al. RNA-guided human genome engineering via Cas9. Science. 339 (6121), 823-826 (2013).