sRNAs that repress transcription have been engineered to create orthogonal and composable regulators that can be used to construct RNA-only transcriptional networks

techniques, such as transposition with Mu 11, Tn5 12 and Tn7 13, have been used to add fluorescent gene markers to plasmids

Alongside this, we used pKJK5::gfp, a plasmid lacking these additional payload genes

none of the organisms identified are known to produce particularly small cells that could pass 0.2 µm filters

Nanopore sequencing is used to rapidly and inexpensively assess part composition and create a composition-to-barcode index

For the terminus, we used primers that amplified part of the dcp gene: 5′-TTGAGCTGCGCCTCATCAAG-3′ and 5′-TCAACGTGCGAGCGATGAAT-3′

Capra Biosciences harnesses the superpowers of a biofilm-forming organism Marinobacter atlanticus to make fat-soluble products like retinol which is used in anti-aging face creams, and high-end industrial lubricants for electric car engines

Some startups, like MicroByre, have been working to domesticate those reluctant, recalcitrant, and rebellious microbes by adapting them for use in the lab and creating genetic tools to manipulate them

Filter and trim

binding of the Mob protein to the 52-bp sequence could thus allow the formation of a protein-DNA complex with a double function: relaxosome formation andmob gene regulation.

Student's t-Test

3.3.1 Citation Styles

focus on the content

approaches, which typically encapsulate bacteria in hydrogels, have produced deployable optical sensors for explosives14, heavy metals15 and chemical inducers16,17

current state-of-the-art whole-cell biosensors for field applications34,35, such as hydrogel-based20,36 and liquid-in-a-cartridge devices37

Translocation to a cell’s surface utilizes a signal peptide (for inner membrane translocation) and AIDAc as an outer membrane autotransporter pore

AHL communication modules with functional devices built from the biological components of the las18, tra18, rpa18, rhl19, cin19 and esa20 quorum-sensing systems

Receiver plasmids contained the synthetic PlasR promoter fused to a strong RBS (BBa_B0034) and GFP gene (BBa_E0040) from the IGEM registry. These plasmids additionally contained a pMB1 origin from pET28a and either a chloramphenicol (CmR) or a kanamycin (KanR) selectable marker.

folA gene from the evolved and naive populations was amplified by PCR

Evolution in regulatory regions rapidly compensates the cost of nonoptimal codon usage

masked by mechanisms affecting the abundance and function of the protein products of the transferred genes downstream to transcription and translation steps, and, therefore, being unrelated to codon bias

very high antibiotic concentrations used in this study (up to 30-fold the minimal inhibitory concentration [MIC]) potentially minimized the effect of sequence composition on fitness

Computational studies that analyzed HGT events among bacterial genomes revealed that HGT frequency positively and strongly correlates with the similarity of tRNA pools between donors and acceptors

5′-end mRNA, the fitness contribution of mRNA folding stability dominates over that of codon optimality.

We constructed an Escherichia coli donor strain that can deliver a genetic payload into target recipients by broad-host-range bacterial conjugation

The mixes were spotted on PBS + 1.5% agar plates and incubated at 37 °C

In contrast, it is the removal of rare taxa that would appear to remove valid data

programming environmental microbes have been applied to a wide range of organisms. Some examples of newly-programmable microbes include

Because biosensors detect at the temporal scales of microbial behaviors, they hold the potential to monitor fast chemical transformations in the environment that are hard to observe, often described as cryptic

and AHL controls in this process remain poorly understood.

There has been an ongoing debate on whether rarefaction is a statistically defensible procedure.

instead of being used to endlessly study one environment after another, in situ techniques should be coupled with efforts to model HGT in attached communities by facilitating comparisons between predicted and experimental observations, and assist in establishing predictive frameworks.

Only fluorescence genes have been used for the in situ detection of HGT in natural environments, as no substrate has to be added to detect the product of the expressed reporter gene.

The fluorescent marker gene approach is of specific importance in this thesis.Compared to the other reporter gene approaches it allows for quasi-immediate detection of plasmid transfer, even in individual cells, without theneed of substrate addition or taking the strains out of their natural environ-ment.

Christensen et al. (5) which used a specific positive induction of gfp in the recipient rather than a down regulation in the donor as in the system used here.

The mobilizing, conjugal plasmid can, now, after retromobilization, co-occurring with RSF1010 in the transconjugant, be subsequently isolated within its original environmental host.

mobilizable plasmids might less frequently reach dead ends once acquired, since they can utilize the conjugal connections build through adapted resident plasmids in their new host

To date, serine integrases have not been used extensively in plant systems

Prokaryotic genes are often subjected to frequent horizontal gene transfer [12–16], and one therefore might expect that this could also affect their gene connectivity.

In many cases, such variations reflect differences in the essentiality of genes. Genes with higher connectivity are often more essential for the reproductive success of a cell or organism

our newly developed biosensor our studies demonstrate the detection of DPO in rodent and human samples.

Bacteria evolve rapidly not only by mutation and rapid multiplication, but also by transfer of DNA, which can result in strains with beneficial mutations from more than one parent.

conditions amenable to optical outputs for fluorescent reporters, they cannot be applied in native communities growing within non-transparent environmental materials

Seedlings were transferred to 10-cm square petri dishes containing Fahraeus agar (Somasegaran and Hoben, 1994) covered with sterile filter paper

While flowFrame objects store the underlying data matrix inthe exprs slot as an R object, cytoframe objects store the matrix (as well as the data from theother slots) in a C data structure that is accessed through an external pointer. This allows for greateroptimization of data operations including I/O, parsing, transformation, and gating.

Collaborative publications, in which the student is one of several authors/creators, are permitted. In every case, the Preface must clearly describe the student's contribution to the research and creation—including, where applicable, the student's role in publications with several authors, or in material created by several authors.

Students enrolled in DSRT 999 are not eligible for the doctoral medical insurance subsidy unless they are otherwise registered as a full-time student.

The dashed lines show the threshold at which cells are considered to be in the ON state (102 for all switches)

cell pellet suspended in 20 μl of Lyse and Go PCR Reagent (Thermo Scientific)

or both rows and columns ("free")?

at the 100% threshold, microbial diversity could be overestimated by as much as 156.5% when using the full-length gene

was digested with PstI (TaKaRa, Japan) to prevent the copy number from being underestimated (66).

meritocracy is imperfect. The best and brightest do not always win. Butthe idea that meritocracy is nothing but a myth is demonstrably false, indeed absurd

For science to succeed, it must strive for the nonideological pursuit of objectivetruth.

That the absurd comparison of the COVID19 crisis to genocide made it intoprint is consistent with a growing body of evidence suggesting that, in the “right” circles,one can make almost any ridiculous claim, as long as one frames it as advancing “SocialJustice.”

These pieces fail to acknowledge the progress that has been made and continuesto be made toward equality, fairness, and justice throughout the Western world

typically without citation to supportingevidence

But examples of a few overlooked individualsdo not imply that meritbased selection is ineffective—

Specifically, he mentions cutoffs thatwould treat 50–75% of applicants as “qualified,” which stops short of abandoning meritaltogether.

The existence of objective reality, for example,which CSJ denies

Again, though, the identity or positionality of the mentor is irrelevant tothe evaluation of merit when using these sorts of quantitative metrics.

But the value of just counting, however imperfect, should be obvious

The first isthat their rejection of objectivity undermines their credibility. If there is no objectivity, thentheir claims are not objectively true.

These perspectives often view science as a tool of power, are hostile to the centralliberal principle of free inquiry and open discussion, and are closed to calls to justify theirclaims on scientific grounds

These core principles, which have served us well for centuries, are under attack byideologies originating in postmodernism and Critical Theory,6,10,14 versions of which rejectobjective reality in favor of “multiple narratives” promulgated by different identity groupsand “alternative ways of knowing.”

Because objectivetruth exists, ultimate consensus among truthseeking actors—scientists—is possible.

we offer a liberal, humanisticalternative that is compatible with maximizing scientific advances.

Thestatuses, identities, and demographics of scientists are irrelevant to this great sifting ofvalid versus invalid ideas.

Fulfilling this responsibility, however, is being hindered by a new, alarming clashbetween liberal epistemology and identitybased ideologies.

How did we get here? Science provided solutions to such calamities as famineand plague, transforming them “from incomprehensible and uncontrollable forces ofnature into manageable challenges.”

guides(color=guide_legend(nrow=2, byrow=TRUE))

Don’t waste time and energy collecting metrics you’ll never be able to use. Learn as much as you can from the failures themselves, and then move on.

Many people rush to collect data without first considering whether they can come up with a model powerful enough to interpret the data. I

Analysis of your incident data may shed light on your incidents. But incident data really can’t tell you anything about your reliability.

the diversity of native plants and animals declined with the invasion of alien plants, communities and ecosystems were rarely negatively affected

an alien species is invasive only if it threatens the native biodiversity of recipient ecosystems and negatively affects ecosystem functions, which could lead to detrimental socioeconomic impacts

most known alien microorganisms are pathogens51, their impact is expected to be negative at all levels.

Several invasive microorganisms infect mainly native plants as their primary host

transfer efficiencies between them were similar to those seen with parental partners (0.21)

The resulting constructs (the pMATING series) could be self-transferred through a variety of prokaryotic and eukaryotic recipients employing such a rationally designed conjugal delivery device.

Assuming each sequence in our downloaded database represented a unique species

the antagonistic symbiotic relationship between the wasps and their insect hosts allowed the wasp larvae to acquire insect pathogens that subsequently evolved to benefit the wasp

labs(fill='Legend Title')

Earn 75,000 bonus miles when you spend $4,000 on purchases in the first 3 months from account opening, equal to $750 in travel.

Bins used for plotting:

amperometric sensor was then modified to detect an endocrine disruptor.

optical signals are often difficult to detect in situ5,6,7,8

Some people argue that the Welch’s t-test should be the default choice for comparing the means of two independent groups since it performs better than the Student’s t-test when sample sizes and variances are unequal between groups, and it gives identical results when sample sizes are variances are equal. In practice, when you are comparing the means of two groups it’s unlikely that the standard deviations for each group will be identical. This makes it a good idea to just always use Welch’s t-test, so that you don’t have to make any assumptions about equal variances.

Non-parametric tests don’t make as many assumptions about the data, and are useful when one or more of the common statistical assumptions are violated. However, the inferences they make aren’t as strong as with parametric tests.

In practice, a data set with a sufficient number of replicates forms an approximately normal distribution

Transcription of the araBAD genes under control of the pBAD promoter is induced about 300-fold above a basal uninduced level within 3 s of the addition of arabinose to cells growing on glycerol in minimal salts medium

This system can efficiently create genome modifications in multiple phages without the need for counterselection

Nonparametric statistics are often preferred to parametric tests when the sample size is small and the data are skewed or contain outliers.

when to use parametric versus nonparametric tests [8,9,10]

Noteworthy for its longstanding influence is thebook “Nonparametric statistics for the behavioralsciences” by Siege

Vmax has a low-level natural resistance to kanamycin but is more susceptible to other antibiotics. When selecting on Kan plates, Vmax with KanR will simply grow faster than Vmax without KanR. Larger colonies=KanR. Other antibiotics select against Vmax but must have a dosage change compared to E. coli.

Wash the culture in fresh LB before adding glycerol, to avoid problems with reviving the culture later

ecovery media (5 BHI + v2 + 68 mM Sucrose )

Four inducible promoters were found to function in V. natriegens

Vmax@SGIDNA.com

To prepare V. natriegens cells for −80 °C storage, an overnight culture of V. natriegens was washed in fresh medium before storage in glycerol

For best results, use only LB-Miller agar plates. Do not use LB-Lennox or LB-Luriaplates (Vmax™ growth will be suboptimal).

Vibrio natriegens strain containing a major extracellular nuclease knockout and insertion of an IPTG-inducible T7 RNA polymerase cassette for expression of genes under a T7 promoter.

(a)colE1(b)SC101(c)RSF1010

we successfully demonstrated challenging clonings such as Gibson Assembly with 5 and Aquacloning with 3 fragments as well as 8 part golden-gate reactions.

all downstream manipulations with V. natriegens ATCC14048 Δdns cells were performed with antibiotic selection (kanamycin (200 μg/mL), chloramphenicol (2 μg/mL), carbenicillin (100 μg/mL))

A survey of 230 diverse bacterial and archaeal genomes found evidence of DNA methylation in 93% of genomes, with a diverse array of methylated motifs (834 distinct motifs; average of three motifs per organism)

conjugative plasmids have broad-host ranges23, are resistant to restriction-modification systems24, are easy to engineer with large coding capacities25, and do not require a cellular receptor26 that would provide a facile mechanism for bacterial resistance.

Bacterial cells are typically one thousandth the volume of mammalian cells, which places them near the edge of instrument detection. At this size it can be challenging to differentiate viable cells from debris of similar size

Detailed descriptions, assumptions, limitations and test cases of many popular statistical methods for ecological research can be found in the GUSTAME server (Buttigieg and Ramette, 2014), and in the review by Paliy and Shankar (2016).

condensing the information into two- or three-dimensional spaces. A very good overview of techniques to achieve this was written by Paliy and Shankar (2016).

There are several widely used tool collections, e.g., QIIME 2 [13], mothur [14], usearch [15], and vsearch [16], and 1-stop pipelines, e.g., LotuS [17], with new approaches continually being developed, e.g., OCToPUS [18] and PEMA [19]

Recently, redox-responsive biomolecules such as phenazines have been used in several electrochemical strategies to interrogate a range of biological activities30,31 and to control gene expression in living cells32,33, where the redox status of the biomolecules could be measured or manipulated by application of electronic potentials

There are two main reasons to use logarithmic scales in charts and graphs.

One reason for choosing a dot plot rather than a bar chart is that it is less cluttered. We will be learning other benefits of dot plots in this and future posts.

Using a logarithmic axes on a bar graph rarely make sense.

1000 cells per sample were sorted into microwells by flow cytometry, heat treated to extract DNA, and used as template for a duplex ddPCR reaction

The genre kicked off with “Maps of Time” (2004), by David Christian, and includes such practitioners as Mr. Harari, Steven Pinker, Jared Diamond and Francis Fukuyama.

geNomad uses marker information to demarcate provirus boundaries.

here's a trivial example of why the "standard" geometric mean  is not a good estimate of central tendency when you have values near  zero.

o improve accuracy, Sony hasintroduced the Weighted Least Square Method (WLSM) al-gorithm. In WLSM, the square value of residuals are individu-ally weighted

Impossible Foods, Zymergen, Pivot Bio, Novartis, and Calyxt

and for P. putida, Ap, Sill" Su, or Nal at 200 j.tg/ml, Cm at400 j.tg/ml, and Km or Tc at 25 j.tg/ml

Ramsay, J. P. & Firth, N. Diverse mobilization strategies facilitate transfer of non-conjugative mobile genetic elements. Curr. Opin. Microbiol. 38, 1–9 (2017)

A cryptic rolling-circle plasmid from a commensal Escherichia coli has two inversely oriented oriTs and is mobilised by a B/O plasmid

RIVET has been used as a promoter-trap method to identify bacterial and fungal genes induced during infection (6–8) and as a tool to analyze the spatiotemporal patterns of induction of such genes (9).

Package ‘flowWorkspace’

rbind2(cytoset, cytoset)

taxa should be monophyletic

glyphosate (the most common pesticide used globally) inhibits growth of bacteria

Studying the effects of temperature—along with other climate change-induced environmental disruptions

thepotential "glycosyl transferase" enzymes thathave been chose

docked to theactive site of the glycosyltransferase enzyme by usingAutodock software

However, its hightoxicity restricts its use in the pharmaceutical field

Glycosyltransferaseenzymes catalyse the glycosylation reactions, which leads to a decreasein the toxicity of celastrol.

E. coli BL21(DE3) strain was preferred forthe production of recombinant protein inthis study, thanks to all these properties andits well-known knowledge

This molecule,unlike other VOCs, originated only from lung cancer cells. It has been found that theHeneicosane molecule works in nature as a pheromone of some species.

Celastrol is one of the most important triterpenoid molecules used in ancient times in China

A vector with the mScarlet-I protein construct under the nptII promoter from

conjugation of the TOL plasmid was found to increase with the specific growth rate of both the donor and recipient (Smets et al., 1993, Seoane et al., 2010). This suggests a relationship of faster-growing bacteria (r-strategists) being able to transfer and receive degradative plasmids better than the slower-growing K-strategists

For each culture, a total of 106 cells were added. Naphthalene was added to the NAH7 conjugation cultures by adding naphthalene crystals to the CBM medium which ensured the medium was continually saturated with naphthalene

Novosphingobium aromaticivorans F199donor harboring pNL1; 16S copy number: 3

All data were analyzed using RStudio, version 1.4.1103. Experimental replicates for all data were averaged and reported with standard error.

Bars are colored by whether naphthalene was added and at which hour the sample was taken.

To quantify conjugation via fluorescence-activated cell sorting (FACS), genes encoding constitutive fluorescence of mScarlet-I were inserted onto the NAH7 and pNL1 plasmids.

these transfer regimens were evident for not only the donor transferring the plasmid, but also the recipient receiving it (Sysoeva et al., 2019).

Historically, bacterial evolution has favored two general ecological growth strategies: high reproduction rate (r-strategists or copiotrophs) or optimal resource utilization (K-strategists or oligotrophs)(Atlas and Bartha, 1997)

environmentally-friendly

Care should be taken to avoid severe ecological disturbance when introducing an alien species.

confirmed to participate in protein remineralization in sediments

Abundance and diversity of archaea in coastal ecosystems

Archaea mediate four major processes that strongly impact the coastal blue carbon ecosystems, namely, CO2 fixation, organic biopolymer transformation, CH4 production, and CH4 oxidation

That's greater than taking all the humans who lived throughout time, multiplied by the number of grains of sand on Earth, multiplied by the number of atoms in the universe.

Digestion is recommended whenever DNA input is greater than 75 ng

DNA fragmentation by restriction digestion prior to dropletgeneration enables optimal accuracy by separating tandemgene copies, reducing sample viscosity, and improving templateaccessibility for input samples >66 ng per well

we show that they have shaped the evolution and host range of plasmids

We found a hierarchy of avoidance, with plasmid genes avoiding 6-bp palindromes significantly more on average than core and non-core chromosomal genes

For the most common target length (6 bp) we show that target avoidance is strongly correlated with the taxonomic distribution of R-M systems and is greater in plasmid genes

As an initial proxy for restriction targets, we first analysed the avoidance of short palindromes in each pangenome component for k=4 and k=6

bacteria possessing cognate R-M systems have higher rates of horizontal gene flow between them

This addictive quality may contribute to their occasional occurrence on MGEs such as plasmids: around 10.5% of plasmids carry R-M systems (Oliveira, Touchon, and Rocha 2014) and experiments have shown R-M system carriage can lead to increased plasmid stability in cells

We report that the EcoRI restriction endonuclease-modification methylase (rm) gene pair stabilizes plasmids that carry it

maths historian George Joseph’s book The Crest of the Peacock: Non-European Roots of Mathematics (1991) and the ongoing encyclopaedic series Science Across Cultures, edited by Helaine Selin

Artificial proteins fine-tuned to five distinct lysozyme families showed similar catalytic efficiencies as natural lysozymes, with sequence identity to natural proteins as low as 31.4%. ProGen is readily adapted to diverse

requires experiments that alter climate in combination with manipulations of microbial communities.

While measurements at each scale do exist for plant-associated microbes, integration across scales is missing for any single system

Data that combine multiplicatively, like rates, are actually very common outside of economics too. The key is to recognize when a measured variables is affected by many (semi) independent forces, each of which scales that variable up or down — rather than simply adding or subtracting a fixed amount to it. This is often true in the natural sciences.

the use of constructed wetlands to generate cellulosic biofuel using waste nitrogen from wastewater treatment

total abundances of archaea and bacteria in these sediments increase with latitude

Increases in solar radiation, temperature and freshwater inputs to surface waters strengthen ocean stratification and consequently reduce transport of nutrients from deep water to surface waters, which reduces primary productivity

received 3-CA every day (press-disturbed), every 2, 3, 4, 5, 6, or 7 days (intermediately-disturbed), or never (undisturbed)