L4.3 Uses of Plasmids Flashcards
general use of plasmid
- manipulate gene expression
advantageous characteristics of plasmids
-flexible
-versatile
-safety
-cost-effective
Types of plasmids based on function
- Cloning plasmid- insert genes in the organism
- Replication plasmid - carry AMR, degradative, express functions
- Gene-knockdown
- Reporter Plasmids
- Viral Plasmids
- Genome-engineering plasmids
food vectors, drug delivery, hormone production of the cells, sources of horizontal gene transfer
Other functions of plasmids
-plasmids 101 or proteins
-GFP and luciferase
- chemical level in an environment
-produce enzymes for controlled changes
- produce synthetic viruses
is a process wherein the
genetic information of an organism is
changed resulting in a mutation. Occurs
spontaneously in nature or as a result
of exposure to mutagens
Mutagenesis
Difference bet in vivo and in vitro
In vivo - inside the organism
In vitro - induction or spontaneous mutation in the lab
Methods of in vitro mutagenesis
random
site directed
deletion
insertion
Techniques to produce protein with better properties
- substrate binding and substrate conversion
-change in thermal and pH stability
-substrate binding site increase specificity - resistance to protease
-alter allosteric regulation
a methods used to generate DNA sequences with mutated codes, insertions or deletions
site-directed mutagenesis
Site-directed mutation is called
point mutation
conformational structure did not change
Because of the artificial nature of
plasmids, they are commonly referred
to as
vectors
Examples of restriction genes
EcoR1 - E. coli {cleaves bet G and A)
Sac1- S. achromogenes
are enzymes that
cleave DNA at specific nucleotide sequences.
The sequence recognized is often _____
nucleotides long.
Restriction endonucleases
four to six
two fragments of DNA are joined together by
DNA ligase
are enzymes that
cleave DNA at specific nucleotide sequences.
The sequence recognized is often four to six
nucleotides long.
Gene cloning
Importance of gene cloning
pest resistnace
bioremediation
heart attack theraphy
human growth hormone
Prcoedures in gene cloning
step 1
- Chromosomal DNA vectior and isolation
-Vector DNA (plasmid, viral vectors)
- Vector replicates
Prcoedures in gene cloning
step 2
insetion of GOI
- yield a recombinant vector having the GOI
