Week 7: Biotechnology and Bioengineering

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49 Terms

1
Genetic Engineering
The modification and manipulation of an organism's genome using technology, began in the early 1970s
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2
Herbert Boyer and Stanley Cohen
Created the first GMO in 1973 by inserting antibiotic resistance genes into E. coli
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3
Genentech
Produced the first commercial GMO product (human insulin) in 1978 using recombinant E. coli
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4
Genetic Vectors
DNA molecules used to carry foreign genetic material into host organisms
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5
Plasmids
Primarily used to transform bacteria, some (shuttle vectors) work in eukaryotes
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6
Bacteriophage

Derived from λ and M13 genomes, used to transduce bacteria, Can integrate (lysogenic) or not integrate (lytic) into the host genome

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7
Cosmids
Hybrid of plasmid and λ phage DNA, used to transform E. coli
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8
Artificial Chromosomes
Circular (BAC) or linear (YAC) vectors for large insert sizes
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9
BAC (Bacterial Artificial Chromosome)
Can carry inserts up to 100–300 kb
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10
YAC (Yeast Artificial Chromosome)
Can carry inserts up to 1–2 Mb
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11
Plasmid Vectors
Small, circular, double-stranded DNA molecules engineered to carry, replicate, and express foreign genetic material
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12
Key Components of Plasmid Vectors

Origin of replication, multiple cloning site, promoter/regulatory elements, selectable marker, and reporter genes

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13

Origin of replication

Allows the plasmid to replicate independently within the host cell

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14

Multiple Cloning Site

Contains unique sequences for inserting foreign DNA

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15

Promoter/Regulatory Elements

Drive the transcription of inserted genes

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16

Selectable Marker

Antibiotic resistance genes to select for positive cells

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17

Reporter Genes

Visual markers like lacZ (β-galactosidase) or GFP (green fluorescent protein)

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18
Restriction Enzyme Cloning
A technique for generating recombinant DNA using restriction enzymes to cut DNA and DNA ligase to join fragments
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19
Sticky Ends
Single-stranded DNA overhangs created by restriction enzymes, used for ligation
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20
Type IIS Restriction Enzyme
Generates user-defined sticky ends for Golden Gate Assembly
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21
Golden Gate Assembly
A restriction enzyme-based technique for assembling multiple DNA fragments into a single construct
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22
DNA Ligase
Joins compatible sticky ends of adjacent DNA fragments
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23
Gibson Assembly
A cloning technique that joins linear vector and DNA inserts into a circular DNA molecule without restriction enzymes
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24
5’ Exonuclease
Chews back the 5’ ends of DNA fragments in Gibson Assembly
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25
DNA Polymerase
Fills gaps by extending the 3’ ends in Gibson Assembly
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26
DNA Delivery Methods
Techniques for introducing DNA into host cells
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27
Transformation
DNA delivery into prokaryotic cells, methods include heat shock and electroporation
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28
Transfection
DNA delivery into eukaryotic cells, methods include lipofection, gene gun, and microinjection
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29
Heat Shock Transformation
A method of transformation using calcium chloride and heat to make cells competent
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30
Electroporation
Uses electric pulses to create temporary membrane pores for DNA entry
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31
Lipofection
DNA encapsulated in cationic lipid vesicles (liposomes) for delivery into cells
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32
Gene Gun
DNA-coated gold particles propelled into cells, common in plants
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33
Microinjection
DNA injection using a fine needle, used for oocytes or embryos
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34
Viral Vectors

Engineered viruses used to deliver genetic material into plant and animal cells

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35
Lentivirus
Integrates into the host genome, effective for dividing and non-dividing cells
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36
Adenovirus
Does not integrate into the genome, leads to transient expression
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Adeno-associated Virus
Provides stable but non-integrating gene expression
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38
Retrovirus
Integrates into the host genome of dividing cells
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39
Protein Engineering
The design and modification of proteins to enhance or alter their function, stability, or specificity
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40
Directed Evolution
A technique that mimics natural selection to improve or modify protein function
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41
Random Mutagenesis
Introduces random mutations via error-prone PCR or DNA shuffling
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42
Screening/Selection
Identifies improved protein variants from a library of mutants
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43
Rational Design
An engineering approach involving knowledge-based modifications to improve or alter protein function
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44
Site-directed Mutagenesis
Introduces specific mutations to improve protein function
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45
De Novo Design
The computational and experimental creation of entirely new proteins from scratch
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46
Computational Design
Uses AI and molecular modeling to predict sequences for stable, functional 3D structures
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47
CRISPR/Cas
A gene-editing technology derived from a bacterial immune defense mechanism
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48
Cas9
A protein that forms a complex with guide RNA (sgRNA) to cleave complementary DNA sequences
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49
sgRNA
Small guide RNA that directs Cas9 to a specific DNA sequence for cleavage
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