A visual continuous spatial directed evolution method is disclosed. The host grows and moves in a solid culture space, the host carrying a foreign target gene to be evolved and containing a gene element that assists the evolution of the target gene, the target gene being correlated with the growth and movement of the host. Depending on different spatial distribution patterns formed in the solid culture space during the growth and movement of the host, screening is performed to obtain an evolved product. This method is carried out directly in the solid culture space. Depending on images of different spatial distribution morphologies visible to the naked eye that are locally formed, selection of evolved products is performed without the need for liquid fed-batch culture equipment. In addition, the evolution effect is visually observed through the infection spots formed during evolution, so that no real-time monitoring equipment is required.

The disclosure provides amino acid sequence variants of Bacillus thuringiensis (Bt) toxins and methods of producing the same. Some aspects of this disclosure provide methods for generating Bt toxin variants by continuous directed evolution. Some aspects of this disclosure provide compositions and methods for pest control using the disclosed variant Bt toxins.

Disclosed herein are methods for the production of low to medium expressing constitutive promoters in bacteria and promoters produced therewith.

Provided herein are methods and compositions for screening using an in vitro transcription/translation (TXTL) system. In some embodiments, a screening method can include: providing a library of genetic candidates to be screened, each genetic candidate encoding one or more protein of interest; expressing each genetic candidate in an in vitro transcription/translation (TXTL) system, thereby producing a plurality of compositions each comprising the one or more protein of interest; and subjecting the plurality of compositions to an assay for assessing a desired phenotype, thereby determining whether the one or more protein of interest exhibits the desired phenotype.

The present disclosure relates to methods to evolve macromolecular machines and improved macromolecular machines identified and made by the methods. In some embodiments, the improved macromolecular machines include improved tethered ribosomes. Also disclosed are systems and methods for making and using the improved tethered ribosomes.

The present disclosure relates to methods to evolve macromolecular machines and improved macromolecular machines identified and made by the methods. In some embodiments, the improved macromolecular machines include improved tethered ribosomes. Also disclosed are systems and methods for making and using the improved tethered ribosomes.

Disclosed are methods for identifying bio-molecules with desired properties (or which are most suitable for a round of directed evolution) from complex bio-molecule libraries or sets of such libraries. Some embodiments of the present disclosure provide methods for virtually screening proteins for beneficial properties. Some embodiments of the present disclosure provide methods for virtually screening enzymes for desired activity and/or selectivity for catalytic reactions involving particular substrates. Some embodiments combine screening and directed evolution to design and develop proteins and enzymes having desired properties. Systems and computer program products implementing the methods are also provided.

The present disclosure provides methods of generating recombinant bacteriophage genomes. Specifically, the present technology provides methods of integrating a heterologous nucleic acid sequence into a linear bacteriophage DNA genome, and isolating recombinant bacteriophages that express the heterologous nucleic acid sequence.

Described is a process for producing a mutant bacterium which exhibits improved survival and/or growth under a selected growth condition, the process comprising the steps of: (a) generating a pool of mutant bacteria by transposon mutagenesis with an activating transposon (TnA), wherein the TnA comprises a promoter capable of increasing transcription of a gene at or near its insertion site; (b) growing bacteria from the mutant pool under the selected growth condition and under one or more reference conditions to produce two or more test cultures; and (c) comparing the distribution of TnA insertions between test cultures to identify a first class of genes which are disadvantageous for growth and/or survival under the selected growth condition and a second class of genes which are advantageous for growth and/or survival under the selected growth condition.

The disclosure provides amino acid sequence variants of Bacillus thuringiensis (Bt) toxins and methods of producing the same. Some aspects of this disclosure provide methods for generating Bt toxin variants by continuous directed evolution. Some aspects of this disclosure provide compositions and methods for pest control using the disclosed variant Bt toxins.