rigid pili, such as Inc N, P and W, conjugate at rates 2–4 orders of magnitude higher on solid surfaces

flexible pili (e.g. Inc F, H and I), transfer equally well in liquid as in solid surfaces

synthetic arrays of regulatory RNA could be created that achieved our design goals

these commonly used artifacts are small code snippets that are entirely functional in nature and, therefore, when used in isolation, don't enjoy copyright protection at all.

The donor and recipient populations grow according to the following standard exponential growth equations

The primary color palette uses three colors. Grey and white should be employed liberally ashigh-contrast elements. Red should be used sparingly to label and decorate symbols and information.We use this pink/red because it is more gender-neutral than the typically masucline color palettes ofother space/geospatial-focused groups and it is distinct from the green/earth tones of conservationgroups. It’s also effective for drawing attention to specific elements

apparent permissiveness (AP). It is defined as the ratio of the relative abundance of an ASV in the transconjugant pool to the corresponding recipient pool

triggering homologous recombination and creating genetic instability10,11,12,13

We applied machine learning to explain how specific interactions controlled the promoters’ transcription rates.

Wilcoxon rank sum tests were used to test for significant differences between two groups

Each template molecule in a sample is tagged with a UMI sequence consisting of 10–20 random bases, which can subsequently be used to sort and analyze reads based on their original template molecule

Three distinct rRNA sequences exist in the operon, encoding for the two subunits of the ribosome; the 5S and 23S sequences code for the large subunit, while the 16S sequence encodes for the small subunit.

demonstrate that targeting of 16S variable regions with short-read sequencing platforms cannot achieve the taxonomic resolution afforded by sequencing the entire (~1500 bp) gene

New methods are needed that can manage and help organize this scale of data. To address this, we consider the general problem of computing an approximate distance between two sequences

we inferred the parameters of a linear function fit to the log transformed gDNA concentration versus detection time (Fig. 2d–g). The inferred slope of the linear function is determined by the cell doubling time (~0.5 h) and intercept is determined by the background mutation frequency

gDNA concentration versus the detection time for the (d) EC sensor, (e) ST sensor, (f) SA sensor and (g) CD sensor

A threshold of GFP fluorescence 400 arbitrary units (a.u.) was used to determine the detection time for each gDNA concentration.

expressed from the IPTG-inducible promoter Phyperspank

cell-based DNA detection is relatively simple and cost-effective. In comparison, other DNA detection methods such as next-generation sequencing and quantitative polymerase chain reaction require specialized instruments

Next-generation engineered bacteria hold tremendous promise

supplemented with 0.3 mM diaminopimelic acid (DAP)

p + theme(legend.position="top")

ubstantial contributions to conception anddesign, or acquisition of data, or analysis andinterpretation of data;

Your questions should be reasonably scoped. If you can imagine an entire book that answers your question, you’re asking too much.

A method was developed for oriented immobilization of bacteriophage T4 through introduction of specific binding ligands into the phage head using a phage display technique

Several review papers on PFAS occurrence, distribution, and exposure have been published in the last decades.

UMAP embedding of Record-seq data

metabolites and RNA are short-lived, omics-based measurements of transient stimuli only yield a snapshot of highly dynamic processes.

Lactococcal strains were routinely grown in M17 medium (49) supplemented with 1% glucose (GM17 medium) or, when needed, in chemically defined SA medium (26) at 30°C without shaking

Acinetobacter baylyi ADP1 (30) and ADP1-ISx (11) were grown at 30°C in LB-Miller (10 g NaCl, 10 g tryptone and 5 g yeast extract per liter) or ABMS minimal medium (40)

We have sought to investigate the combined impacts of complex substrates and presence of electron acceptors.

a biologically driven cryptic cycle was determined by identifying metabolically active sulfate reducing and sulfur oxidizing lineages co-locating within the sediments, effectively masking sulfide production through re-oxidation back to sulfate.

Why Make Memories?

Corynebacteriumglutamicum

Making universal media for bacterial strains

typically between 1010 and 1011 microbial cells per wet-weight gram of faeces7,8,9.

This figure is commonly mentioned in the microbiome literature, but its source has been difficult to ascertain

increased activity has brought with it a degree of hype and misinformation, which can undermine progress and public confidence in the research

Compare and Merge revisions √ Learn about comparing and merging documents

Proteinase K to the reaction to a final concentration of 50–100μg/ml. Incubate at 37–56°C for at least 1 hour. R

Differences in growth rate, e.g. induced by sub-lethal antibiotic concentrations or temperature, can also significantly bias conjugation rate estimates.

The default (50nM) works well with the standard protocol used at the Whitehead/MIT Center for Genome Research

a rerun of those identical amplicons (from the same reaction tubes) on a different date yielded “normal” results

A normal blood sodium level is between 135 and 145 milliequivalents per liter (mEq/L).

adjust the sample richness of each ecosystem by adding to it an estimate of the number of unobserved species

Adjusting for sample size when comparing different groups of observations without discarding data is widely prevalent in the sciences, and discarding data to adjust for unequal sample sizes is the exception

there is substantial negative bias in the estimates

since estimates for alpha diversity metrics are heavily biased when taxa are unobserved, comparing alpha diversity using either raw or rarefied data should not be undertaken

We currently do not account for measurement error in microbial diversity studies.

unique property of microbiome experiments and alpha diversity analysis is that samples do not faithfully represent the entire microbial community under study

The samples are not of particular interest, except that they reflect the environment from which they were sampled

Based on these results and well-established statistical theory, we advocate that investigators avoid rarefying altogether. We have provided microbiome-specific extensions to these tools in the R package, phyloseq.

Well-established statistical theory is available that simultaneously accounts for library size differences and biological variability using an appropriate mixture model

since the most commonly  used models are underdispersed relative to the data, standard errors are almost always zero. The result is that ecologists see incredibly small p values, but replicating results is rare.

We care about producing high-quality software and making our papers reproducible.

the key aspect of a protocol’s ability to accurately reflect the original sample composition can only be assessed with the help of standardized controls like mock communities

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

You cannot claim a credit for education expenses paid with tax-free funds. You must reduce the amount of expenses paid with tax-free grants, scholarships and fellowships and other tax-free education help.

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)