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- W974733802 abstract "During investigation, the function of deoxyribonucleic acid (DNA) sequences must indirectly demonstrate expression by means of the existence of messenger ribonucleic acid (mRNA) by ribonuclease protection, in situ hybridization, or in situ polymerase chain reaction (PCR). The next step consists of cloning the DNA in an expression vector—a plasmid that, in addition to the usual plasmid components such as the bacterial replication origin and antibiotic resistance, contains a gene with a promoter or a coding area. Depending on whether the experimenters want to examine a promoter or a complementary DNA (cDNA), they replace the region of the DNA that interests them. A vector is chosen with an easily demonstrable reporter gene, before which the experimenter can insert the promoter. Enhancer sequences can also be explored in this manner by using a vector that contains a basic promoter and whose activity in the cloning of regulatory elements can be upregulated or downregulated. When determining the section in which the DNA activity can be found, mutagenesis can be used to remove sequences. The choice of a proper reporter gene is decisive in this kind of investigation. When working with cDNA, experimenters have a multitude of expression vectors available to them that deliver a promoter and consequently wait to be associated with an open-reading pattern. Viral promoters such as that of the cytomegalovirus or from simian virus 40 permit a quite intense expression in mammalian cells, and functional investigations can be frequently accomplished immediately. Cell-typical promoters permit expression based on certain tissue limitations, such as in the manufacture of transgenic animals, and controllable promoters already exist." @default.
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- W974733802 date "2007-01-01" @default.
- W974733802 modified "2023-09-25" @default.
- W974733802 title "Investigating the Function of DNA Sequences" @default.
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