17.1: Recombinant DNA
- Page ID
- 74593
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- 17.1.2: Introduction of recombinant DNA into cell and replication- Vectors
- Vectors used to move DNA between species, or from the lab bench into a living cell, must meet three requirements: (1) They must be autonomously replicating DNA molecules in the host cell. (2) They must contain a selectable marker so cells containing the recombinant DNA can be distinguished from those that do not. (3) They must have an insertion site to accommodate foreign DNA. Usually a unique restriction cleavage site in a nonessential region of the vector DNA.
- 17.1.4: Make and Screen a cDNA Library
- The first step in making a cDNA library is to isolate cellular mRNA. This mRNA extract should represent all of the transcripts in the cells at the time of isolation, or the cell’s transcriptome. This term is used by analogy to genome. However, a genome is all of the genetic information of an organism. In contrast, a transcriptome (usually eukaryotic) reflects all of the genes expressed in a given cell type at a moment in time.