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Expression vectors produce proteins through the transcription of the vector's insert followed by translation of the mRNA produced, they therefore require more components than the simpler transcription-only vectors. Expression in different host organism would require different elements, although they share similar requirements, for example a ...
For example, prokaryotes expression vectors would have a Shine-Dalgarno sequence at its translation initiation site for the binding of ribosomes, while eukaryotes expression vectors would contain the Kozak consensus sequence. The promoter initiates the transcription and is therefore the point of control for the expression of the cloned gene.
Usually the ultimate aim of expression cloning is to produce large quantities of specific proteins.To this end, a bacterial expression clone may include a ribosome binding site (Shine-Dalgarno sequence) to enhance translation of the gene of interest's mRNA, a transcription termination sequence, or, in eukaryotes, specific sequences to promote the post-translational modification of the protein ...
Non-lytic insect cell expression is an alternative to the lytic baculovirus expression system. In non-lytic expression, vectors are transiently or stably transfected into the chromosomal DNA of insect cells for subsequent gene expression. [23] [24] This is followed by selection and screening of recombinant clones. [25]
Regulation of gene expression, or gene regulation, [1] includes a wide range of mechanisms that are used by cells to increase or decrease the production of specific gene products (protein or RNA). Sophisticated programs of gene expression are widely observed in biology, for example to trigger developmental pathways, respond to environmental ...
An expression cassette is a distinct component of vector DNA consisting of a gene and regulatory sequence to be expressed by a transfected cell. [1] In each successful transformation, the expression cassette directs the cell's machinery to make RNA and protein(s). Some expression cassettes are designed for modular cloning of protein-encoding ...
BAC vectors contain a replicon derived from E. coli F factor, which ensures they are maintained at one copy per cell. [4] Once an insert is ligated into a BAC, the BAC is introduced into recombination deficient strains of E. coli by electroporation. Most BAC vectors contain a gene for antibiotic resistance and also a positive selection marker. [2]
For most prokaryotic chromosomes, the replicon is the entire chromosome. One notable exception comes from archaea , where two Sulfolobus species have been shown to contain three replicons. Examples of bacterial species that have been found to possess multiple replicons include Rhodobacter sphaeroides (two), Vibrio cholerae , [ 3 ] and ...