Bio3400 Chapter 19 Recombinant DNA Technology
  1. A              enzyme binds to DNA at a specific recognition sequence and cleaves the DNA to produce restriction fragments.

  2. Most recognition sequences are              , and restriction enzymes can cleave these sequences in an offset manner to produce         ends.

  3. DNA fragments can be         using restriction enzymes and          such as plasmids and then replicated in host cells.

  4. Other          that can carry larger target DNA include phages, cosmids, and            artificial chromosomes (BACs).

  5. Often the cloned gene of interest should be             into proteins.             vectors are engineered to express large quantities of the encoded protein.

  6. Eukaryotic hosts such as        cells are useful to produce eukaryotic proteins since they can perform                    modifications and carry very large DNA fragments.

  7. Bacterial plasmids can be used to transfer genes to plant cells in a process caled                 .

  8. YACs can be used to transfer genes to animal cells in a process caled direct            .

  9. The             chain reaction (PCR) can be used to amplify small quantities of target DNA in vitro.

  10. Genes from whole chromosomes can be cloned to form genomic            . The individual chromosomes can be isolated by       cytometry and pulsed-field gel electrophoresis.

  11. Libraries of transcriptionally active genes in particular cells can be constructed from                DNA (cDNA).

  12. Specific clones can be recovered from a library by              with a probe followed by                  .

  13. Specific DNA sequences can be identified with a           blot and autoradiography.

  14. DNA fragments can be sequenced by          chain termination. Large-scale genome sequencing can be automated by using              dideoxynucleotides.