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Microbiology > Biotechnology > Flashcards

Biotechnology Flashcards

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Biology 101 flashcards
1
Q

Biotechnology

A

The use of microorganisms, cells, or cell components to make a product (foods, antibiotics, vitamins, emzymes )

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2
Q

Recombinant DNA technology (rDNA)

A

The insertion or modification of genes to produce desired proteins

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3
Q

Clone

A

Population of genetically identical cells arising from one cell; each carries the vector

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4
Q

Selection

A

Selecting for a naturally occuring microbe that produces a desired product

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5
Q

Mutation

A

Mutagens cause mutations that might result in a microbe with a desirable trait

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6
Q

Site-directed mutagenesis

A

A targeted and specific change in a gene

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7
Q

Restriction enzymes

A
  • Cut specific sequences of DNA
  • Destroy bacteriophage DNA in bacterial cells
  • Methylated cytosines in bacteria protect their own DNA from digestion
  • create blunt ends or staggered cuts known as sticky ends
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8
Q

Vectors

A
  • Carry new DNA to desired cells
  • Must be able to self-replicate (ori-origin of replication)
  • Plasmids and viruses can be used as vectors
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9
Q

Shuttle vector

A

Exist in several different species and can move cloned sequences among various organisms

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10
Q

What is the Polymerase Chain Reaction (PCR)

A

The process of increasing small quantities (amplifying) of DNA for analysis

  • Used for diagnostic tests for genetic diseases and detecting pathogens
  • Reverse transcription PCR uses mRNA as template
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11
Q

What are the steps of the polymerase chain reaction -PCR

A
  1. Incubate target bacteria at 94 dc for 1 min; this temp will separate the strands (hydrogen bonds break)
  2. Add primers, nucleotides, and DNA polymerase
  3. Incubate at 60 dc for 1 min; this allows primers to attach to single-stranded DNA
  4. Incubate at 72 dc for 1 min; DNA polymerase copies the target DNA at this temperature
    Second cycle- repeat the cycle of heating and cooling to make two more copies of target DNA
    *Denature-Anneal-Extension
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12
Q

Transformation

A

Cells take up DNA from the surrounding environment

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13
Q

Electroporation

A

Electrical current forms pores in cell membranes

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14
Q

Protoplast fusion

A

Removing cell walls from two bacteria; allows them to fuse

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15
Q

Gene gun

A

Most frequently used on plant cells to insert foreign DNA

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16
Q

Microinjection

A

Small needle used to inject DNA (mostly animal cells)

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17
Q

What are genomic libraries?

A

Collections of clones containing different DNA fragments

  • An organism’s DNA is digested and spliced into plasmid or phage vectors and introduced into bacteria
  • At least one clone exists for every gene in the organism
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18
Q

What is cDNA?

A

Complementary DNA is made from mRNA by reverse transcriptase

  • Used for obtaining eukaryotic genes because eukaryotic DNA has introns that do not code for protein.
  • mRNA has the introns removed, coding only for the protein product
19
Q

Blue-white screening for selecting clones

A

Uses plasmid vector containing ampicillin resistance gene (amp^R) and B-galactosidase gene lacZ

  • Bacteria is grown in media containing ampicillin and x-gal, a substrate for B-galactosidase.
  • Only the the bacteria that picked up the plasmid will grow in the presence of ampicillin/ Bacteria that hydrolyze x-gal produce galactose and in indigo compound. The indigo turns the colonies blue; The bacteria that cannot hydrolyze x-gal produce white colonies
20
Q

Colony hybridization for selecting clones

A

Use DNA probes (short segments of single-stranded DNA complementary to the desired gene.

21
Q

List the advantages and disadvantages of using E. coli for genetic engineering

A

Advantages- Easily grown and its genomics are known

Disadvantages- Produces endotoxins and does not secrete its protein products

22
Q

List the advantages and disadvantages of using Saccharomyces cerevisiae (eukaryotic yeast) for genetic engineering

A
  • Easily grown and has a larger genome than bacteria

- Expresses eukaryotic genes easily

23
Q

List the advantages and disadvantages of using Plant cells and whole plants for genetic engineering

A
  • Express eukaryotic genes easily

- Plants are easily grown, large-scale, and low cost

24
Q

List the advantages and disadvantages of using mammalian cells for genetic engineering

A

Advantages-Express eukaryotic genes easily
-Can make products for medical use
Disadvantages- Harder to grow

25
Q

Subunit vaccines

A

Made from pathogen proteins in genetically modified yeasts

26
Q

DNA vaccines

A

-Non pathogenic viruses carrying genes for pathogen’s antigens (Not approved for human use)

27
Q

Gene therapy

A

To replace defective or missing genes

28
Q

Gene silencing (SiRNA)

A

Small interfering RNAs (SiRNAs) bind to mRNA, which is then destroyed by RNA-induced silencing complex (RISC)

29
Q

Gene silencing process

A
  1. An abnormal gene, cancer gene, or virus gene is transcribed in a host cell
  2. SiRNA binds mRNA
  3. RISC breaks down the RNA complex
  4. No protein expression occurs
30
Q

RNA interference (RNAi)

A

Inserts DNA encoding siRNA into a plasmid and transferred into a cell

31
Q

Shotgun sequencing

A

Sequences small pieces of genomes which are assembled by a computer

32
Q

Metagenomics

A

The study of genetic material directly from environmental samples (i.e. soil)

33
Q

The Human Genome Project

A

Sequenced the entire human genome

34
Q

The Human Proteome Project

A

Will map proteins expressed in human cells

35
Q

Bioinformatics

A

Understanding gene function via computer- assisted analysis

36
Q

Proteomics

A

Determining proteins expressed in a cell

37
Q

Reverse genetics

A

Discovering gene function from a genetic sequence

38
Q

Southern blotting

A

DNA probes detect specific DNA in fragments (RFLPs) separated by gel electrophoresis

39
Q

Northern blotting

A

DNA probes used to identify RNA

40
Q

DNA fingerprinting

A

Used to identify pathogens-PCR microarrays and DNA chips can screen samples for multiple pathogens
-Differs from medicine because it requires proper collection of evidence and establishing a chain of custody

41
Q

Nanotechnology

A

Bacteria can make molecule-sized particles

  • Nanospheres used in drug targeting and delivery
  • Lipid nanoparticles used to deliver the mRNA in the moderna and Pfizer covid-19 vaccines
42
Q

Ti Plasmid

A

Occurs in Agrobacterium tumefaciens

  • Integrates into the plant genome and causes a tumor-like growth
  • Can be used to introduce rDNA into a plant
43
Q

What are some of the agricultural applications of rDNA

A 
B+ toxin
Herbicide resistance
Suppression of genes-antisense DNA
Nutrition
Human proteins
44
Q

Identify some of the safety and ethical issues using DNA technology

A
  • Need to avoid accidental release into the environment
  • Genetically modified crops must be safe for consumption and for the environment
  • Who will have access to an individual’s genetic information?

Microbiology (7 decks)

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