Chapter 9 Flashcards by Lani Manners

biotechnology

the use of microorganisms, cells, or cell components to make a product

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example of products biotechnology

foods, antibiotics, vitamins, enzymes

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recombinant DNA (rDNA) technology

the insertion or modification of genes to produce desired proteins

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vector

self replicating DNA molecule used to transport foregin DNA into a cell

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cells with vectors multiply to form

clone

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clone

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

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selection

selecting for a naturally occurring microbe that produces a desired product

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mutation

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

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site directed mutagenesis

a targeted and specific change in a gene

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restriction enzyme cut specific sequences of

DNA

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restriction enzymes _______ phosphodiester bonds between individual nucleotides at a specific recognition sequences on both strands DNA, and they are able to cut or digest the DNA sequence the same way every time

hydrolyze

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restriction enzymes can occur

naturally

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restriction enzymes typically cleave

unmethylated DNA

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restriction enzymes create blunt ends or staggered cuts known as

sticky ends

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___ ________ attaches the complementary sticky ends to one another this produces recombinant DNA molecule

DNA ligase

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vectors carry new DNA to

desired cells

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vectors must be able to

self replicate

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______ and ______ can be used as vectors

plasmids, viruses

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shuttle vectors

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

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PCR

polymerase chain reaction

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PCR is a process of increasing small quants of DNA for

analysis

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PCR is used for diagnostic tests for

genetic diseases and detecting pathogens

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reverse transcription PCR uses ____ as a template and reverses the normal flow of genetic information. Utilizing the enzyme _______ ________ mRNA is used to create cDNA

mRNA, reverse transcriptase

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what is cDNA

complementary DNA

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the steps of PCR

1. denaturation 2. priming 3. extension

denaturation PCR step is what temp

high temp

denaturation PCR results

DNA is incubated at high temp which causes the strands tp separate by breaking the hydrogen bonds

priming PCR temp

low temp

priming PCR results

DNA is incubated at a relatively low allowing primers to attach to the single stranded target DNA

extension PCR temp

intermediate temp

extension PCR results

DNA is incubated at an intermediate temp at which the DNA polymerase rapidly replicates DNA

PCR occurs in a machine called

thermocycler

PCR components

DNA polymerase, DNA primers, dATP, dCTP, dTTP, DGTP,

inserting foregin DNA into cells 3 types

transformation, electroporation, protoplast fusion

transformation

cells take up DNA from the surrounding environment

electroporation

electrical current forms pores in cell membranes

protoplast fusion

removing cell walls from 2 bacteria allows them to fuse

DNA can be inserted into a cell by 2 ways

Gene gun, microinjection

gene gun is used in

plants

microinjection is a technique that uses a _____ _______ with a diameter much smaller than the cell, and is able to puncture the plasma membrane, so that DNA can be injected through it

glass micropipette

genomic libraries are collections of clones containing different

DNA fragments

an organisms DNA is digested and spliced into _______ or _______ ______ and introduced into bacteria

plasmid, phage vectors

complementary DNA is made from mRNA by

reverse transcriptase

genomic libraries are used for obtaining eukaryotic genes because

eukaryotic DNA has introns that do not code for protein

mRNA has the _______ removed, coding only for the protein product

introns

synthetic DNA builds genes using a

DNA synthesis machine

in cloning it is necessary to select the particular cell that contains your specific ___ of interest

gene

blue white screening uses _______ _____ containing ________ resistance gene and b-galactosidase

plasmid vector, ampicillin

for blue white screening bacteria is grown in media containing ampicillin and X gal, a substrate for b- galactosidase, only the bacteria that picked up the plasmid will

grow

white colonies

bacteria that picked up the the recombinant plasmid will not hydrolyze lactose

blue colonies

bacteria with the intact lacZ gene will hydrolyze the x-gal and produce blue colonies

colony hybridization

common method of identifying cells that carry specific gene

colony hybridization uses DNA probes, what are these?

short segments of single stranded DNA complementary to the desired gene

e coli advantages for using it in making a gene product

easily grown and its genomics are unknown

e coli disadvantages for using it to make a gene product

produces endotoxins and does not secrete protein products

saccharomyces cereviase is what

bakers yeast

saccharomyces cereviase - easily grown and has a ________ genome than bacteria - expresses __________ genes easily - likely to continuously secrete ________

larger, eukaryotic, product

plant cels and whole plants - express ________ genes easily - plants are easily grown, large scale and ___ cost

eukaryotic, low

mammalian cells - express _________ genes easily - can make products for ______ use - ______ to grow

eukaryotic, medical, harder

subunit vaccines

made from pathogen proteins in genetically modified yeasts

DNA vaccines usually _______ plasmid including a gene coding for a ____ protein

circular, viral

gene therapy

to replace defective or missing genes

gene editing uses

CRISPR

gene editing

to correct genetic mutations at specific locations

2 ways to conduct gene silencing

small interfering RNAs (siRNAs) RNA interference (RNAi)

small interfering RNAs (siRNAs)

bind to mRNA which is then destroyed by RNA induced silencing complex

RNA interference (RNAi)

inserts DNA endowing siRNA into a plasmid and transferred into a cell

shotgun sequencing

sequences small pieces of genomes which are assembled by a computer

metagenomics

the study of genetic material directly from environmental samples

Human Genome Project

sequences the entire human genome

Human Proteome Project

will map proteins expressed in human cells

bioinformatics

understanding gene function via computer assisted analysis

proteomics

determining proteins expressed in a cell

reverse genetics

discovering gene function from a genetic sequence

southern blotting DNA probes detect specific DNA fragments ________ which are restriction fragment length polymorphisms separated by ___ ________

RFLPs (rif lips), gel electrophoresis

genetic testing

screening of parental or fetal tissue for several hundred possible genetic diseases

DNA fingerprinting

is used to identify pathogens

PCR microarrays and DNA chips

can screen samples for multiple pathogens

forensic microbiology can be used to

find proper collection of evidence and establish a chain of custody

nanotechnology deals with the design and manufacture of extremely small electronic _______ and mechanical devices built at the ________ level of matter

devices, molecular

bacteria can make nanoparticles from

gold, silver, selenium and cadmium

Ti plasmid occurs in

agrobacterium tumefacines

Ti plasmid is a plasmid that can be used to introduce recombinant DNA into a

plant

Ti plasmid integrates into the plant genome and causes

tumor like growth

Bt toxins is normally produced by

Bacillus thuringiensis

Bt toxins can be produced at larger amounts by genetically modified

Pseudomonas fluorescens

Bt toxin is used as

pest control for insects that feed on crop plants

suppression of gene agricultural example

tomatoes that stay firm after harvest because the enzyme that breaks down pectin is suppressed

suppression of genes is achieves with

antisense DNA

antisense DNA is a length of DNA complementary to the ____ for the enzyme is synthesize and then is taken up by the cell where it binds to mRNA which inhibits _______ then the DNA-RNA hybrid is degraded by ______ in the cell and this frees up the antisense DNA to disable another mRNA

mRNA, translation, enzymes

when using DNA technology we need to avoid accidental release into the

environment

when making genetically modified crops we must make sure they are safe for

consumption and environment