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Biotechniques

Mizzou, Spring 2015, Biochem Final

TermDefinition
human Chinese scientists genetically modify _____ embryos with the CRISPER/Cas9 technology
genetic engineering nucleic acid biotechnology is modifying the genetic materials of living cells so that they will produce new substances or perform new functions; this is also called ______ ______
gel electrophoresis a separation technique where the separation of DNA based on its LENGTH, and the separation occurs inside a gel within an electric field; because DNA is negative, it will migrate to the positive electrode
restriction endonuclease recognize specific DNA sequences and break the phosphodiester bond in both strands (i.e. they cut DNA)
palindromes recognition sequences are often _______ (the left to right sequence on one strand is the same as right to left sequence on the complementary strand)
sticky ends digested DNA generates _____ _____, which are over-hangs of single stranded DNA
complementary compatible sticky ends can re-join because they are _________
bacteriophages these protect bacteria against infection (viruses)
methylating bacteria prevent digestion of their own DNA by the restriction enzymes by ________ nucleotides in the restriction enzyme recognition sequence
recombinant ________ DNA is the central part of genetic engineering; covalently linking DNA fragments from more than one source
ligase recombinant DNA was first created in vitro using restriction endonuclease and DNA ________
restriction use ________ enzymes to create recombinant DNA
recombinant DNA is formed by fist cutting DNA from two sources to create complementary sticky ends, second the complementary ends are joined by base-pairing between overhanging single-stranded regions, and finally DNA ligase creates covalent bonds between strands
vectors self-replicating forms of DNA
cloning once inserted into the vectors, large amount of the gene can be replicated to allow the study of this gene; the process of making identical copies
plasmid circular DNA that can self-replicate in bacteria
bacteriophage a virus that infects and replicates bacteria
adenovirus a virus that infects mammalian cells
retrovirus a virus that infects mammalian cells
cloning the insertion of foreign DNA into the DNA of these vectors allows replication of the vector and foreign DNA together in the host cell
plasmid extra-chromosomal circular molecules of DNA capable of replication inside bacteria; they are used to clone foreign DNA, often containing genes
DNA ligase plasmid DNA and foreign DNA are joined by ____ _____
ori origin of replication
selection gene usually an antibiotic resistance gene to allow growth of only the bacteria containing the plasmid
polylinker the region of DNA containing restriction sites for inserting foreign DNA to be studied
transformed the plasmid containing the inserted DNA is "_______" into E. coli bacteria which replicates the plasmid
cloned as the bacteria replicated, many copies of the foreign gene are produced, or ________
expression vectors certain plasmids are constructed to allow transcription and translation of a foreign gene in bacteria, these are called ____ ____
introns genes to be expressed by expression vectors must contain only the exons without introns because bacteria will not remove _____ from mRNA
transgenic organism an organism containing a heritable foreign gene or TRANSGENE
reverse transcription _______ ______ uses reverse transcriptase from a retrovirus to produce a cDNA molecule from mRNA; avoids intron problem this way
somatic changes change in genetic makeup of specific tissues that are NOT inherited by offspring
germline changes genetic changes made in cells that form reproductive cell (egg & sperm); these are INHERITED by future progeny; not theatrical or legal in human subjects
gene therapy ex) severe combined immune deficiency syndrome (SCIDS) patients have mutations in adenosine deaminase (ADA) that affects the T cell immune function; these patients undergo ____ ____ to hope to place a good copy of the ADA gene in the bone marrow
randomly DNA is injected into a fertilized mouse egg & _____ integrates into the genome (nowadays the transgenic can be targeted)
polymerase chain reactions (PCR), how scientists are able to detect DNA in very small quantities (e.g. at crime scenes)
replicating PCR is a technique for ______ specific fragments of DNA in vitro
hours PCR allows a single DNA molecule to be specifically replicated into billions of copies in a few hours
thermophiles a type of bacteria that is able to grow under extreme heat conditions (80-105 degrees C)
thermo aquaticus ______ ______ bacteria live in the hot springs of Yellowstone National Park & contain a heat-stable DNA polymerase (Taq polymerase)
Taq polymerase PCR requires ____ ______, a heat stable DNA polymerase
exponential PCR amplification of DNA is _______ (i.e. the target DNA duplicates every cycle)
PCR applications of ____ include: identifying pathogenic organisms, genetic disease diagnosis, sex determination of embryos, and a LOT MORE
sgRNA CRISPER (clustered regularly interspaced short palindromic repeats)/Cas system relies on a single guide RNA (_____) to direct the Cas9 protein
Created by: oliviarees95