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Hello Microtwjc community!

This week we are going to have a discussion on a recent article I have published with Laura Bowater (@Lauramcbow) and Paul Hoskisson (@PaulHoskisson) on engaging students on STEM degrees with the Antimicrobial Resistance issue. Yes this is not something we have discussed before as we have generally focussed on research papers. However, all of us at some point do interact with students at all levels. We are also interested to see what you all have to say on this subject….

You can find the paper here:

Inspiring STEM undergraduates to tackle the AMR crisis.

Things to discuss:

  1. Can you think of other ways to engage students on this topic?
  2. Do you have specific examples that would also work as case examples?
  3. Can we improve our engagement of under and post-graduates on topical issues within the learning environment?

See you all on twitter at 8 PM Tomorrow night!


I have chosen a paper based on a technique I saw being exploited at a recent conference I attended. The paper can be found using the link below and I hope it really is open access:

Long-Lived Intracellular Single-Molecule Fluorescence Using Electroporated Molecules

This is a bit different from our usual papers but there are aspects of this technique that make one think of what we can now do! However the usual rules apply:

Discussion points:

1. Is the paper well written and concise?

2. Are the experiments well designed?

3. Anything you would have done differently?

Abstract: Studies of biomolecules in vivo are crucial to understand their function in a natural, biological context. One powerful approach involves fusing molecules of interest to fluorescent proteins to study their expression, localization, and action; however, the scope of such studies would be increased considerably by using organic fluorophores, which are smaller and more photostable than their fluorescent protein counterparts. Here, we describe a straightforward, versatile, and high-throughput method to internalize DNA fragments and proteins labeled with organic fluorophores into live Escherichia coli by employing electroporation. We studied the copy numbers, diffusion profiles, and structure of internalized molecules at the single-molecule level in vivo, and were able to extend single-molecule observation times by two orders of magnitude compared to green fluorescent protein, allowing continuous monitoring of molecular processes occurring from seconds to minutes. We also exploited the desirable properties of organic fluorophores to perform single-molecule Förster resonance energy transfer measurements in the cytoplasm of live bacteria, both for DNA and proteins. Finally, we demonstrate internalization of labeled proteins and DNA into yeastSaccharomyces cerevisiae, a model eukaryotic system. Our method should broaden the range of biological questions addressable in microbes by single-molecule fluorescence.

We have a paper for this weeks MicroTWJC number 48…

Global Population Structure and Evolution of Bordetella pertussis and Their Relationship with Vaccination

The papers own significance statement is below. This is an interesting study with an important implication that should lead to a good discussion. See you all on Tues 6th May at 8 PM (British Time)

IMPORTANCE Whooping cough is mainly caused by Bordetella pertussis, and current vaccines are targeted against this organism. Recently, there have been increasing outbreaks of whooping cough, even where vaccine coverage is high. Analysis of the genomes of 343 B. pertussis isolates from around the world over the last 100 years suggests that the organism has emerged within the last 500 years, consistent with historical records. We show that global transmission of new strains is very rapid and that the worldwide population of B. pertussis is evolving in response to vaccine introduction, potentially enabling vaccine escape.

A small challenge for this week: Can you read this paper without the need to refer to the supplementary data?

UPDATE: You can find the storify of our conversation here –> Storify of #MicroTWJC number 48



For this weeks journal discussion I have decided to return to Salmonella. The paper of choice has the wonderful title of:

Genome and Transcriptome Adaptation Accompanying Emergence of the Definitive Type 2 Host-Restricted Salmonella enterica Serovar Typhimurium PathovarmBio vol. 4 no. 5 e00565-13

I have chosen this paper not only because it ends up focussing on bacterial motility and that it is Salmonella, one of the bugs I use to study motility in the lab! The title seems a bit misleading but lets read the paper before we make any comments. Following a trend of papers we have covered, this study utilises genome analysis to question species variability/adaptation. I also feel it is apt that we go back to discussing a genome paper this week with the recent announcement that Fred Sanger died. What better why to take note of the work he did by discussing a paper from one of the cohorts of microbiologists based at the Sanger Centre in the UK. I have also reused the discussion questions from week #35 as they fit with this paper


  1. Is the paper well written, easy to understand and thoroughly explained?
  2. Are the methods appropriate?
  3. What do the results tell us – how significant is the impact of this research?
  4. What research can and should this follow on to?

So people here is next week’s paper for discussion.

Microanatomy at Cellular Resolution and Spatial Order of Physiological Differentiation in a Bacterial Biofilm

I had a hard choice and I have decided on one of two stories (the other one was on phages) that caught my attention at the BACNET13 meeting I attended in Poland recently.

The wonderful images produced by the Hengge lab while dissecting the role of c-di-GMP in biofilm formation won. You will need to use the online material as well as the paper for this one.

We have discussed c-di-GMP and Biofilms sometime back and it is a subject that started my own journey into the fascinating world of Caulobacter crescentus as a young Post-Doc. Regine Hengge gave an excellent talk on this story at BACNET. The field of c-di-GMP itself played a central role at this meeting. What was clear to me is that the field has begun to move on from the biochemical characterisation of the components involved to returning to the underlying question of what c-di-GMP is regulating, why and how? Clearly exciting times are ahead for the field.

As per usual our discussion will focus on:

1) Did the paper read well?

2) Could you understand the regulatory network?

3) Would you have done anything differently?

4) where next?