Quiz 5 Flashcards by Zach Moreno

T Cell Receptors (TCRs)

Bind antigenic peptides that have been processed and presented with the major histocompatibility complex (MHC) of an antigen presenting cell

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T Helper Hybridoma KZH

Created by fusing normal helper T cells from immunized mice with a T lymphoma cell

Contain a reporter construct with lacZ gene encoding beta-galactosidase

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Reporters (ex. GFP) are used to monitor changes in ____

gene expression in whole animals, tissues, and individual cells

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Beta-galactosidase produced by activation of the IL-2 promoter is released from cells using ____

TritonX100 detergent

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Chromogenic substrate turning from yellow to red when hydrolyzed by beta-galactosidase

Chlorophenol red galactoside (basis of color change)

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Antigen used in 11.B

Lysozyme

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Antigen-presenting mouse B cell hybridoma, use MHC Class II to present antigen to KZH TCR

LK35.2

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A laminar flow hood is used when working with ____

mammalian cell cultures

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A CO₂ incubator is used to keep gas levels at ____% and maintain optimal pH for mammalian cell culture of ____

5, 7.4

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CD molecule most likely to be expressed by T cells used in 11.B

CD4 (Helper T cells)

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Antibiotics

Chemicals produced by microorganisms that inhibit particular steps in metabolic processes

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Narrow-spectrum Antibiotics

Effective only against a limited number of pathogens (ex. Azithromycin, Clindamycin, Erythromycin)

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Broad-spectrum Antibiotics

Attack many different kinds of bacteria (ex. Amoxicillin, Tetracycline, Quinalones)

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Cidal

Antibiotics that kill microorganisms

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Static

Antibiotics that only inhibit growth

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Beta-lactams (from Penicillium, Cephalosporium)

Inhibit synthesis of bacterial cell wall peptidoglycan

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Macrolides (from Streptomyces erythreus)

Inhibit rRNA associated with the 50S ribosome

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Aminoglycosides (from S. griseus, S. fradiae, Micromonospora purpurea)

Inhibit 30S ribosome function

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Tetracyclines (from S. aureofaciens)

Inhibit bindings of aminoacyl t-RNA to ribosomes (blocks peptide chain formation)

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Penicillium cross streak plate results

Of the four plated bacteria, only E. coli was able to grow at all on the plate

Since penicillin is a beta-lactam inhibiting peptidoglycan synthesis, all other species (Gram+) were inhibited

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Kirby-Bauer Method

Uses standardized inoculum (MH or TSA plates), and high potency antibiotic discs to measure the inhibition zone in mm

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Minimal Inhibitory Concentration (MIC)

The lowest concentration of antibiotic that will prevent the growth of a particular microorganism, provides a measure of susceptibility

McFarland Standards

Used to standardize the bacterial inoculum used for antibiotic sensitivity tests, using a black line to compare solution turbidity which is related to bacteria concentration

Antibiotics used in 12.B

Erythromycin, Penicillin, Tetracycline, Gentamicin

Bacteria used in 12.B

*E. coli* and *S. aureus*

A microorganism is most sensitive to an antibiotic in the ____ phase

exponential

Microbial resistance mechanisms to penicillin

Gram- species unaffected by beta-lactams, Gram+ species replace the terminal D-alanine

Microbial resistance mechanisms to tetracycline

Multidrug efflux pumps, ribosomal protection, and enzymatic inactivation of antibiotic

Horizontal Gene Transfer

The process by which genes are transferred from one mature, independent organism to another Mechanisms of antibiotic resistance, used in molecular biology and genetic engineering

Conjugation

The process of genetic transfer involving cell-to-cell contact between donor and recipient cells

Transformation

The processes where a piece of free DNA is taken up by a cell and integrated into its genome

Transduction

The transfer of genes between bacterial or archaeal cells by viruses (bacteriophages)

Competent Species

Those that are capable of taking up high MW DNA and recombine it into their chromosomes

Growth on "Mate" Kan+Nal plate in 13 was facilitated by ____

successful transformation of the resistant *Acinetobacter* donor strain genes to the sensitive recipient strain

R Factors

Conjugative plasmids carrying genes for resistance to antibiotics, moves from donor with *tra* gene (encodes sex pilus) and delivered to recipient

Species and plasmid used in 14

*E. coli*, pBBR1MCS-2 plasmid (encodes resistance to kanamycin, *lacZ* gene encoding beta-galactosidase production)

pBBR1MCS-2 Plasmid features:

1. Kanamycin resistance 2. *lacZ* gene 3. Region for conjugation/mobility (*mob*) 4. Multiple cloning site (MCS) - allows insertion of foreign DNA without disruption of plasmid

Restriction sites are recognized by ____

restriction endonucleases (enzymes that cut DNA into pieces)

The recipient strain of *E. coli* in 14 is ____ resistant and ____ sensitive

Nalidixic acid, kanamycin

Sterile X-gal used in 14

A substrate for enzyme beta-galactosidase that hydrolyzes the solution to form an intense blue precipitate (LacZ+ = blue colonies)

Bacteriophages infect host cells first by attaching ____ to a ____

tail fibers, cell receptor

Lytic/Virulent Bacteriophage

Virus nucleic acid converts all cell machinery to manufacture new virus particles

Lysogenic/Temperate Bacteriophage

Viral genetic material comes into equilibrium with that of the host, physically inserts genome to become a **prophage**

Prophage

The latent form of the virus genome that remains within the host but does not destroy it

Phage typing

Specificity of a bacteriophage for a particular host provides the means to detect and differentiate certain pathogenic strains of bacteria by pattern of susceptibility to bacteriophages

The bacteriophage used in lab is a ____ with virulent DNA and a ____ cycle

T phage, lytic

Phage titer

The concentration of infectious phage particles per mL of growth medium (determined by making serial dilutions)

Plaques

Small, clear zones of clearing on otherwise dense lawn of bacterial cells, the result of localized areas of lysis of phage-infected bacteria, generates Plaque forming units (PFUs)

Phage attachment to receptors on surface of bacterial cells is facilitated by the presence of ____

magnesium ions

In 15, chloroform was used to ____

kill all bacteria in bacteriophage/bacterium supernatant

Phages for *Staphylococcus* and *Enterococcus* could be isolated from:

S. = Skin E. = GI tract

Helper T cells (CD4+) read ____ MHC to respond to extracellular infections

class II (aid in helping other immune response cells)

Killer T cells (CD8+) read ____ MHC to respond to intracellular infections in all nucleated cells

class I