Chapter 13: Biotechnology Flashcards
biotechnology
any technological application that uses biological systems, living organisms, or derivatives thereof to make or modify procudts or processes
recombinant DNA
single molecules containing DNA sequences from two or more organisms
what tools are used to make recombinant DNA?
restriction enzymes for xutting DNA into pieces (fragments) that can be manipulated
Gel electrophoresis for the analysis and purification of DNA fragments
DNA ligase for joining DNA fragments together in novel combinations
restriction enzymes
cut double stranded DNA moleules into smaller, noninfectious fragments
recognition sequence or a restriction site
specfic sequence of DNA bases that restriction enzymes recognize
most are 4-6 bp long
make palindromes
some cut so there is a little bit of single stranded DNA
methylases
add methyl groups to cell DNA so that restriction enzymes do not chop up the DNA of the cell they are from becuase they can’t recognixe them
restriction digest
using restriction enzymes in the lab
gel electophoresis
a way to separate DNA fragments
fragments placed in wells on one end of a semisolid gel and an electric field is applied DNA will move toward the positive end, with smaller draments traveling more quickly
What info does gel electophoresis give?
the number of fragments
the size of the fragments
the relative abundance of a fragment
DNA ligase
involved in DNA metabolism
catalyzes the joining of DNA fragments by making phosophodiester bonds between them
joins Okizaki fragments
clone
produce many identical copies
transformation
inserting recombinant DNA into host cells
transfection
insering recombinant DNA into host cells derived from animals
Transgenic
describes a cell with recombinant DNA
True or false: most cells that are exposed to recombinant DNA become transormed with it
false
only a few do
selectable marker genes
ex. genes that confer antibioitic resistabce
often included in recombinant DNA
electroporation
a short electric shock used to create temporary pores in membranes through which DNA can enter
replicon
replication unit
a segement of DNA that contains an origin of replication
How can DNA become part of a replicon?
inserted into host chromsome
can enter host cell as a vector and then integrate or have its own origin of replication
vector
part of a carrier DNA sequence
plasmid
small
one or more restriction enzyme recognition sequence
contain genes that conger resistance
ori and replicate independently
Do most cells take up vectors after interactions?
nope
Do all vector copies have the foreign DNA after ligatio n reactions?
nope
reporter gene
genes whose expression are easily assayed
ec. selectable markers
genomic library
a collection of DNA fragments that comprise the genomw of an organism
cDNA
complementary DNA
cDNA library
snapshot of the transcription pattern of the cell
collections of cDNAs from a particular tissue at a particular time in the life cycle of an organism
synthetic DNA
created from PCR amplifications
knockout experiment
inactivate a gene and see what happens
helps determine the minimal genome
homologous recombination
targets specific genes
insert normal alleles into plasmid and then restriction enxymes insert reports genes in the middle of the normal gene, which prevents it from creating normal proteins
once the recombinant plasmid has been made, it is used to transfect stem cells
swap nonfunctioning allele with functioning in host
stem cell
unspecialized cell that divides and differentiates into specialized cells
antisense RNA
binds by pairing to complementary sense bases on mRNA
ex. microRNA
prevents translation of mRNA
RNA interference
RNAi
a natural mechanism for inhibiting mRNA translation
uses small interfering RNAs which are made in a process similar to the processing od microRNAs (double stranded)
small interfering RNAs
siRNAs
short double stranded RNA that is unwaound to single stranfs by a protein complex that guides one strand to a complementary mRNA and then causes the breakdown of the mRNA
expression vectors
have the characteristics of typical vectors and the extra sequences needed for the foreign gene to be expressed in the host cell
inducible promoter
responds to a specfic signal can be included in an expression vector
tissue-specific promotoer
expressed only in certain tissues at certain times
can be used it localized expression is desired
signal sequences
can be added to expression vectors so the gene product goes where it needs to
pharming
the production of pharmaceuticals in farm animals or plants
advantages of recombinant DNA technology over traditional methods of breeding
-identify specfiic genes
introduce any gene from any organism
ability to generate new organisms quickly
Bioremediation
The use by humans of other organisms to remove contaminants from the environment
