Plasmid vector construction is an essential step for molecular microbiology yet often a time-consuming process. Manipulation of the fungal genome to express genes to activate secondary metabolite production often requires creation of plasmid constructs in a reiterative fashion.

Techniques relying on site-specific or homologous recombination are preferred for construction of large plasmids (>15 kb), while digestion of DNA by restriction enzymes with subsequent ligation of the resulting DNA fragments continues to be the mainstream approach for generation of small- and medium-size plasmids.

A plasmid construct refers to a DNA molecule that has been artificially engineered and assembled into a plasmid vector. It typically contains specific DNA sequences, such as promoter regions, coding sequences, and regulatory elements, which allow the expression of …

In the field of genetic engineering, the most important tool developed was the plasmid vector. This technology has been continuously expanding and undergoing adaptations. Here, we provide a detailed view following the evolution of vectors built throughout the years destined to study microorganisms and their peculiarities, including those whose ...

Plasmid construction involves combining a complete plasmid vector backbone for genetic engineering with a foreign gene of interest, inserting the foreign DNA fragment into the vector. First, a DNA fragment carrying the gene of interest is isolated from its genome, while the plasmid is harvested and purified as described below.

Plasmid vector construction is an essential step for molecular microbiology yet often a time-consuming process. Manipulation of the fungal genome to express genes to activate secondary metabolite production often requires creation of plasmid constructs in a reiterative fashion. Here we introduce a m …

2019年7月23日 · Here we report GT (Guanin/Thymine) standard (GTS) for plasmid construction under which DNA sequences are defined as two types of standard, reusable parts (fragment and barcode). We develop a...

1979年1月1日 · Plasmid vectors provide the recombinant DNA molecule the ability to be introduced into the host cell, to replicate and maintain themselves stably in generations. DNA replication is connected with the cell membrane and is realized by a multienzyme complex.

2012年1月1日 · Here we introduce a modified Quick-change site-directed mutagenesis method that allows for rapid and accurate construction of fungal transformation vectors. Plasmids are self-replicable extrachromosomal circular DNAs ­containing genes of resistance to microbes, degradation of complex organic compounds, and production of toxins and enzymes (1).

2009年1月8日 · Plasmid DNA (pDNA) expression vectors are fundamental to all forms of non-viral gene transfer. In this review, we discuss principles of pDNA design and production including the impact of...