Building synthetic chromosomes from natural DNA.

De novo chromosome synthesis is costly and time-consuming, limiting its use in research and biotechnology. Building synthetic chromosomes from natural components is an unexplored alternative with many potential applications. In this paper, we report CReATiNG (Cloning, Reprogramming, and Assembling Tiled Natural Genomic DNA), a method for constructing synthetic chromosomes from natural components in yeast. CReATiNG entails cloning segments of natural chromosomes and then programmably assembling them into synthetic chromosomes that can replace the native chromosomes in cells. We used CReATiNG to synthetically recombine chromosomes between strains and species, to modify chromosome structure, and to delete many linked, non-adjacent regions totaling 39% of a chromosome. The multiplex deletion experiment revealed that CReATiNG also enables recovery from flaws in synthetic chromosome design via recombination between a synthetic chromosome and its native counterpart. CReATiNG facilitates the application of chromosome synthesis to diverse biological problems.

Complete Genome Sequences of Four Isolated Bacteria from an Undergraduate Microbiology Course Using a Hybrid Assembly Approach.

Three Gram-negative bacteria and one Gram-positive bacterium were isolated from environmental samples in an undergraduate microbiology class on the basis of antibiotic resistance. Isolate DNA was purified, sequenced, and assembled using a hybrid approach. Here, we report the genomes of Acinetobacter johnsonii CSUSB1, Aeromonas hydrophila CSUSB2, Bacillus velezensis CSUSB3, and Comamonas thiooxydans CSUSB4.

Hybrid Assembly of the Quorum-Quenching Isolate Variovorax paradoxus VAI-C Genome Sequence.

Variovorax paradoxus VAI-C was isolated due to its ability to utilize acyl-homoserine lactones (AHLs) as the sole source of carbon, energy, and nitrogen. Here, we present a hybrid assembly of the V. paradoxus VAI-C genome sequence, consisting of a primary chromosome, a secondary chromid, and a plasmid.

Complete Genome Sequences of Pseudomonas alkylphenolica Neo and Variovorax sp. Strain CSUSB, Obtained in Undergraduate Microbiology Courses Using a Hybrid Assembly Approach.

Two Gram-negative bacteria with a high G+C content were isolated from soil in undergraduate microbiology classes by enriching for low nutrient growth and neonicotinoid pesticide tolerance. DNA from these isolates was purified and sequenced using a hybrid approach. Here we report the genome sequences of Pseudomonas alkylphenolica strain Neo and Variovorax sp. strain CSUSB.