polyadenylation

1 زیست شناسی:: پلی‌آدنیله شدن

The vector must have a eukaryotic promoter that drives the transcription of the cloned gene of interest, eukaryotic transcriptional and translational stop sig­ nals, a sequence that enables polyadenylation of the mRNA, and a selectable eukaryotic marker gene (Fig. 3.25). The major features of a eukaryotic expression vector are a eu­ karyotic transcription unit with a pro­ moter (p), a multiple cloning site (MCS) in which to insert a target gene, and a DNA segment with transcription termi­ nation and polyadenylation signals (t); a eukaryotic selectable marker (ESM) gene; an origin of replication that func­ tions in the eukaryotic host cell (orieuk); an origin of replication that functions in E. coli (oriE); and an E. coli selectable marker (e.g. The transfer vector is an E. coli­based plasmid that carries a segment of DNA from AcMNPV (Fig. 3.35A) consisting of the polyhedrin promoter region with flanking upstream AcMNPV DNA, a multiple cloning site, the polyhedrin transcription termination and polyadenylation signal regions with flanking downstream AcMNPV DNA. The promoter sequences that drive expression of the cloned gene(s) and the selectable marker gene(s), and the transcription termination sequences (including polyadenylation signals), must be eukaryotic and are frequently taken from either human viruses or mammalian genes. Strong constitutive promoters and efficient polyadenylation signals are preferred.

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