You are given a plasmid with a gene of interest (an enzyme of your choice), however, the plasmid is unsuitable for downstream processes (recombinant protein expression). Describe how you would: a) Move it into a suitable vector and host. Please name the vector and host and why they were selected. State the appropriate checks to ensure the process is going according to plan. Moving a Gene of Interest into a Suitable Vector and Host

Transferring a Gene Of Interest to a Suitable Vector & Host

Many techniques can be used to insert the gene of your choice into a compatible vector or host. Cloning refers to the process of isolating a gene and inserting it into a vector (plasmid). Then, it’s possible to transfer that gene to a host organism (bacteria or yeast) (Khan et. al. 2020; Sun, et. al. 2020). The typical process starts with extracting the gene desired from an organism. Next, insert it into the vector. The vector can then be transferred to the host cell where the gene is desired and expressed. A common bacterial expression vector pET28a can be used for recombinant expression. After the host and vector have been selected, the gene that is of most interest must be ligated to the vector. Sun et al. (2020): This is a step that requires restriction enzymes to identify and cut the vector at specified recognition sites. By ligation, the gene of interest is then incorporated into a vector. The recombinant gene is then transferred to the host by a suitable technique (conjugation transformation, transfection and so forth) (Chen, 2021). Cont…

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