Starting a new recombinant protein production project in Escherichia coli

Lucero, Dianela Aguilar; Cantoia, Alejo; Ceccarelli, Eduardo A.; Rosano, Germán L.

doi:10.1016/bs.mie.2021.08.019

Search citation statements

Order By: Relevance

Paper Sections

Select...

Citation Types

Supporting

Mentioning

Contrasting

Year Published

2023

2024

Publication Types

Select...

Article2

Book1

Relationship

Self Cite0

Independent3

Authors

Journals

Cited by 3 publications

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

Investigating Protein-Protein Interactions of Autophagy-Involved TNIP1

Samulevich,

Carman,

Aneskievich

2024

Methods in Molecular Biology

View full text Add to dashboard Cite

Investigating Protein-Protein Interactions of Autophagy-Involved TNIP1

Samulevich,

Carman,

Aneskievich

2024

Methods in Molecular Biology

View full text Add to dashboard Cite

Essential factors, advanced strategies, challenges, and approaches involved for efficient expression of recombinant proteins in Escherichia coli

Eskandari,

Nezhad,

Leow

et al. 2024

Arch Microbiol

View full text Add to dashboard Cite

Tips for efficiently maintaining pET expression plasmids

Khananisho,

Cumming,

Kulakova

et al. 2023

Curr Genet

View full text Add to dashboard Cite

AbstractpET expression plasmids are widely used for producing recombinant proteins in Escherichia coli. Selection and maintenance of cells harboring a pET plasmid are possible using either a Tn3.1-type genetic fragment (which encodes a ß-lactamase and confers resistance to ß-lactam antibiotics) or a Tn903.1-type genetic fragment (which encodes an aminoglycoside-3’-phosphotransferase and confers resistance aminoglycoside antibiotics). Herein we have investigated how efficiently pET plasmids are maintained using these two fragments. The study reveals that pET plasmids are efficiently maintained with both Tn3.1 and Tn903.1 genetic fragments prior to the induction of recombinant protein production, and over short induction times (i.e., 2 h). However, over longer induction times (i.e., 20 h), the efficiency of plasmid maintenance depends on the host strain used, and the type of antibiotic selection cassette used. Based on our collective observations, we have 2 general tips for efficiently maintaining pET plasmids during recombinant production experiments. Tip #1: Use a strain with lowered levels of the T7 RNA polymerase, such as C41(DE3). pET plasmids will be efficiently maintained over long induction times with both the Tn3.1 and Tn903.1 genetic fragments, regardless of whether antibiotics are present during cultivation. Tip #2: If a strain with higher levels of T7 RNA polymerase strain is necessary, such as BL21(DE3)), keep induction times short or use a plasmid containing a Tn903.1-type fragment and select with kanamycin.

show abstract

Starting a new recombinant protein production project in Escherichia coli

Cited by 3 publications

References 26 publications

Investigating Protein-Protein Interactions of Autophagy-Involved TNIP1

Investigating Protein-Protein Interactions of Autophagy-Involved TNIP1

Essential factors, advanced strategies, challenges, and approaches involved for efficient expression of recombinant proteins in Escherichia coli

Tips for efficiently maintaining pET expression plasmids

Contact Info

Product

Resources

About