Besides the well-known plasmids, bacteriophages, and transposons, the more recently described integrons can capture multiple gene cassettes, express them, and then integrate them into the chromosome [ 7].
We developed the Promoter-based Gene andembly and Simultaneous Overexpression (PGASO) technique to insert multiple gene cassettes in a predesignated order into the genome of a cell.
If multiple gene cassettes satisfy the criteria for the same region, the one with the highest BLAST score is used as the annotation.
Given that many genes cooperate in various pathways, incorporating multiple gene cassettes in tandem in a transgenic DNA construct for the purpose of genetic modification is often necessary when generating organisms that produce multiple foreign gene products.
Multiple rev/env gene cassettes were obtained from each of seven HIV-1 strains using single genome amplification (SGA) PCR.
The results indicate that integration of the gene cassette into multiple copy loci should be given consideration when designing amylolytic yeast strains.
These include two genes encoding almost identical SpoT (ppGpp hydrolase -like proteins, a gene cassette encoding multiple ATPase domain-containing proteins, and genes for DNA methydrolase -likehelicase.
To create strains deleted for multiple genes, the kanamycin cassette was excised using FLP-mediated recombination, resulting in deletions carrying only a single FRT site (denoted, for example ubiG::FRT in Table 1), as previously described [ 37], with the only difference that the non-selective incubation took place at 37°C and not at 43°C.
Isolating and comparing gene cassette arrays from multiple closely related organisms is necessary to understand their role in natural environments.
Plasmid pCS26 is a low-copy vector that contains a kanamycin resistance gene cassette, and a multiple cloning site (MCS) upstream of a promoterless luciferase operon (luxCDABE) (22).
