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Optimizing fluorescent protein expression for quantitative fluorescence microscopy and spectroscopy using herpes simplex thymidine kinase promoter sequences

Overview of attention for article published in FEBS Open Bio, May 2018
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Mentioned by

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2 tweeters

Citations

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7 Dimensions

Readers on

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28 Mendeley
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Title
Optimizing fluorescent protein expression for quantitative fluorescence microscopy and spectroscopy using herpes simplex thymidine kinase promoter sequences
Published in
FEBS Open Bio, May 2018
DOI 10.1002/2211-5463.12432
Pubmed ID
Authors

Rizwan Ali, Sivaramakrishnan Ramadurai, Frank Barry, Heinz Peter Nasheuer

Abstract

The modulation of expression levels of fluorescent fusion proteins (FFPs) is central for recombinant DNA technologies in modern biology as overexpression of proteins contributes to artifacts in biological experiments. In addition, some microscopy techniques such as fluorescence correlation spectroscopy (FCS) and single-molecule-based techniques are very sensitive to high expression levels of FFPs. To reduce the levels of recombinant protein expression in comparison with the commonly used, very strong CMV promoter, the herpes simplex virus thymidine kinase (TK) gene promoter, and mutants thereof were analyzed. Deletion mutants of the TK promoter were constructed and introduced into the Gateway® system for ectopic expression of enhanced green fluorescent protein (eGFP), monomeric cherry (mCherry), and FFPs containing these FPs. Two promoter constructs, TK2ST and TKTSC, were established, which have optimal low expression levels suitable for FCS studies in U2OS, HeLa CCL2, NIH 3T3, and BALB/c cells. Interestingly, when tested in these four cell lines, promoter constructs having a deletion within TK gene 5'-UTR showed significantly higher protein expression levels than the equivalent constructs lacking this deletion. This suggests that a negative regulatory element is localized within the TK gene 5'-UTR.

Twitter Demographics

The data shown below were collected from the profiles of 2 tweeters who shared this research output. Click here to find out more about how the information was compiled.

Mendeley readers

The data shown below were compiled from readership statistics for 28 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 28 100%

Demographic breakdown

Readers by professional status Count As %
Researcher 10 36%
Student > Bachelor 6 21%
Student > Ph. D. Student 4 14%
Student > Doctoral Student 2 7%
Student > Master 2 7%
Other 1 4%
Unknown 3 11%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 14 50%
Physics and Astronomy 2 7%
Chemistry 2 7%
Agricultural and Biological Sciences 2 7%
Pharmacology, Toxicology and Pharmaceutical Science 1 4%
Other 4 14%
Unknown 3 11%

Attention Score in Context

This research output has an Altmetric Attention Score of 1. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 26 June 2018.
All research outputs
#9,654,166
of 15,171,295 outputs
Outputs from FEBS Open Bio
#282
of 889 outputs
Outputs of similar age
#165,491
of 276,455 outputs
Outputs of similar age from FEBS Open Bio
#12
of 25 outputs
Altmetric has tracked 15,171,295 research outputs across all sources so far. This one is in the 23rd percentile – i.e., 23% of other outputs scored the same or lower than it.
So far Altmetric has tracked 889 research outputs from this source. They receive a mean Attention Score of 2.1. This one has gotten more attention than average, scoring higher than 58% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 276,455 tracked outputs that were published within six weeks on either side of this one in any source. This one is in the 31st percentile – i.e., 31% of its contemporaries scored the same or lower than it.
We're also able to compare this research output to 25 others from the same source and published within six weeks on either side of this one. This one is in the 40th percentile – i.e., 40% of its contemporaries scored the same or lower than it.