Chapter 14, 12 & 7

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Steps in Recombinant DNA technology: 1. [blank_start]Isolate[blank_end] the DNA segment or gene from remaining DNA 2. Use [blank_start]restriction enzymes[blank_end] to cut and join DNA fragments from 2 different sources 3. [blank_start]View[blank_end] DNA fragments 4. Locate DNA fragments with [blank_start]Southern Blotting and Probes[blank_end].

Which is the restriction enzyme that makes staggered cuts in DNA producing sticky ends?

Which enzyme makes straight cuts and produces blunt ends during DNA recombination?

Examples of Recombinant DNA (rDNA) [blank_start]insulin[blank_end] [blank_start]Human growth hormone[blank_end] [blank_start]some vaccines[blank_end] [blank_start]crops with GMO's[blank_end]

What is the purpose of the PCR (Polymerase-Chain reaction)?

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Which enzyme is used during PCR

A [blank_start]southern blot[blank_end] confirms the DNA from a gel is the target DNA. A [blank_start]probe[blank_end] is DNA/RNA with a base sequence complementary to a sequence in the gene of interest. If probes combine at correct spots that confirms the correct [blank_start]target gene[blank_end].

[blank_start]Gene cloning[blank_end] amplifies a specific piece of DNA via a bacteria cell. [blank_start]Vectors[blank_end] determine what is going to transport DNA into a cell. [blank_start]Transformation[blank_end] allows bacteria to take in foreign DNA. [blank_start]Restriction site[blank_end] is the spot to cut plasmid. [blank_start]Ori[blank_end] is the origin of replication. [blank_start]Selectable markers[blank_end] screen for prescence of DNA.

A library consisting only of those DNA sequences that are transcribed into mRNA

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Screening a DNA library

Sanger's Deoxy Sequencing Method

What organisms are unicellular, lack a membrane bound nucleus and membrane bound organelles?

[blank_start]Prototrophic[blank_end]: wild type [blank_start]Auxotropic[blank_end]: mutant type [blank_start]Minimum media[blank_end]: only required by prototrophic bacteria [blank_start]Complete medium[blank_end]: contain all substances required by all bacteria, including auxotrophic

Extra small, circular chromosomal DNA is known as

Gene transfer in bacteria: [blank_start]Conjugation[blank_end]: direct transfer of DNA from on bacterium to another [blank_start]Transformation[blank_end]: bacterium takes up free DNA [blank_start]Transduction[blank_end]: bacterial viruses take DNA from one bacterium to another

[blank_start]Leaderberg and Tatum[blank_end] did a Conjugation experiment: [blank_start]F+ cells[blank_end]: donor cells with the F factor [blank_start]F- cells[blank_end]: recipient cells lacking F factor [blank_start]Sex pilus[blank_end]: connection tube

bacterium with a conjugative plasmid (for example, the F-factor) integrated into its chromosomal DNA. The integration of the plasmid into the cell's chromosome is through homologous recombination.

Antibiotic Resistance [blank_start]Antibiotic resistance[blank_end] comes from the actions of genes located on [blank_start]R plasmids[blank_end] that can be transferred naturally R plasmids have evolved in the past 60 years since the beginning of widespread use of [blank_start]antibiotics[blank_end]. The transfer of R plasmids is not restricted to bacteria of the [blank_start]same or even related[blank_end] species.

A bacterial virus (bacteriophage) carries DNA from one bacterium to another and usually occurs within same or closely related species.

Gene Expression: [blank_start]Constitutive expression[blank_end]: continuously expressed under normal cellular conditions and not regulated [blank_start]Positive Control[blank_end]: stimulate gene expression [blank_start]Negative control[blank_end]: inhibits gene expression [blank_start]Operon[blank_end]: transcriptional unit that contains functionally related genes that include the [blank_start]Promoter[blank_end], [blank_start]Operator[blank_end], and [blank_start]structure genes[blank_end] [blank_start]Regulator gene[blank_end]: not part of the operon, DNA sequence encoding products that affect the operon function, helps control expression of the structural genes of the operon