E. coli is widely used as the host for heterogeneous protein expression for the following advantages: (1) ease of growth and manipulation using simple laboratory equipment; (2) availability of dozens of vectors and host strains that have been developed for maximizing expression; (3) a wealth of knowledge about the genetics and physiology of E ...

There are many types of cloning vectors, but the most commonly used ones are genetically engineered plasmids. Cloning is generally first performed using Escherichia coli, and cloning vectors in E. coli include plasmids, bacteriophages (such as phage λ), cosmids, and bacterial artificial chromosomes (BACs).

1979年1月1日 · This chapter discusses the plasmids of Escherichia coli (E. coli) as cloning vectors. The essence of molecular cloning or recombination in vitro is the joining together in vitro of two or more deoxyribonucleic acid (DNA) fragments. One fragment, called the “vector” or “vehicle,” is capable of replication in some host organism and the ...

2024年11月23日 · A very large number of host organisms and molecular cloning vectors are in use, but the great majority of molecular cloning experiments begin with a laboratory strain of the bacterium E. coli (Escherichia coli) and a plasmid cloning vector.

We demonstrate the usefulness of our design by engineering an E. coli expression vector, pGRASS, where GFP is used as the reporter and allows for the selection of recombinant clones directly on the LB agar plate. In pGRASS, the weak constitutive promoter P lacI q opposes the strong and inducible P T7.

The authors describe proven methods for cloning DNA into plasmid vectors, transforming plasmids into E. coli, and analyzing recombinant clones. They also include protocols for the construction and screening of libraries, as well as specific techniques for specialized cloning vehicles, such as cosmids, bacterial artificial chromosomes, l vectors ...

The desired sequence can be cloned into a binary vector like pBI121, which can replicate in both A. tumefaciens and E. coli [54]. The binary vector contains a smaller T-DNA with the border sequences LB and RB, along with a cloning site and a selectable marker (Figure 14C).

We have therefore developed a novel Escherichia coli cloning vector (termed ‘pJAZZ’ vector) that is maintained as a linear plasmid. Further, it contains transcriptional terminators on both sides of the cloning site to minimize transcriptional interference between vector and insert.

2015年3月16日 · PCR fragments and linear vectors containing overlapping ends are easily assembled into a propagative plasmid by homologous recombination in Escherichia coli. Although this gap-repair cloning approach is straightforward, its existence is virtually unknown to most molecular biologists.

2004年4月5日 · While plasmid vectors were initially designed for gene cloning and DNA analysis in Escherichia coli, shuttle vectors for gene transfer between E. coli and other model organisms for gene function anal...