MicroBio Exam 2

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Cas

(CRISPR-associated) proteins, are endonucleases as defense, and incorporate new spacer regions

(-)ssRNA animal virus influenza virus

-(-) strand, RNA synthetase make complementary RNA strand, which then is the mRNA -RNA synthase also makes (-) strand again from (+) strand as part of replication -Enveloped, pleomorphic (non-uniform), budding virus with segmented genome (separate pieces)

Genomes

-A cell's entire complement of genetic information -Bacterial genomes ca. 1-5 Mbp -Includes genes, regulatory sequences, and no-coding DNA

(+)ssRNA animal virus poliovirus

-Belongs to rhinoviruses, also including coronavirus (+) strand is mRNA -Gets immediately translated when in cytoplasm -A 5'-end guide protein, and 3'-end is poly(A)-tail like on cellular mRNAs -Production of polyproteins that are self-cleaving into individual proteins (like genes of a bacterial operon)

anoxygenic phototrophic

-Carotenoids, e.g., B-carotene -Typically absorb blue light and reflect yellow, red, brown or green

Genome comparison can reveal which of the following?

-Chromosomal islands. -Genes of the core and pan genome. -Regions of putative horizontal gene transfer.

ssDNA bacteriophage ⏀X174

-Contains a circular single-stranded DNA genome inside an icosahedral virion -Contains the (+) strand, so has to make (-) strand/replicative form -From (-) strand mRNA is generated, and more (+) strand(chromosome) to pack into new virions

Basic genetic engineering techniques

-DNA amplification (PCR) -Gel Electrophoresis -Nucleic acid hybridization (probes) -Molecular cloning -Targeted mutagenesis

external influence(mutagenic agents)

-Exposure to radiation -Exposure to chemicals that chemically modify DNA -Infection with an integrating virus

dsDNA animal herpesvirus

-Large group causing different disease in humans, including cold sores, chicken pox, shingles, and infectious mononucleosis -Able to remain latent for extended periods of time (years; reside in nerve cells) -Get re-activated under stress or weakened immune system, can cause nerve damage

dsDNA bacteriophage Mu

-Large virus with an icosahedral head, helical tail, and six tail fibers, genome is linear -Host range is controlled by tail fibers: one type allows infection only of E. coli; other allows infection of other enteric bacteria -Performs lytic and lysogenic pathway -Always integrates: replicated by transposition, at many possible sites of the boat chromosome, via enzyme transposase (brings it to the host)

light reaction needs

-Light-harvesting/antenna pigments in the membrane --Chlorophyll (cyanos) --Bacteriochlorophyll (others) -Light-active molecules have a conjugated electron system -Production of different pigments allows different the kinds phototrophs to absorb different wavelengths of light, while coexisting in the same habitat

Locations for Ammonium- electron donor

-Manure (animal urine) -Produced by bacteria in water and soil as end product of plant and animal waste decomposition -Usually low nontoxic concentrations in soil, air and water and serves an indirect N source for plants -Leaves the system by drainage via groundwater, or, microbial oxidation (nitrification)

What can a genome tell us?

-Metabolic capabilities of an organism -Molecular adaptations to specific environments, e.g., encoding enzymes with features stable at high temperatures -Identify virulence factors -Detecting horizontal gene transfer -Monitoring and diagnosing disease outbreaks

Information on mutations

-Mutations are random changes in ---DNA sequence occurring over time -Most mutations are neutral or deleterious, some are beneficial

Microbial functions in the nitrogen cycle

-Nitrogen has a total of nine oxidation states -Bacteria can perform oxidation or reduction reactions starting at several levels -The ecosystem conditions govern what is possible

Fertilization

-No external electron acceptor -No respiration/membranes involved -No energy conservation with an ion gradient (H+, no proton motive force) -Internal electron cycling, regeneration of NAD+ to achieve redox balance -Substrate-level phosphorylation -Partially oxidized end produced

oxygenic phototrophs

-Phycocyanin absorbs around 550 nm, looks red -Phycocyanin absorbs around 620 nm, looks blue

Why RNA to DNA transition?

-Protect against host ribonucleases (RNAses) -Develop own DNA replication machinery that is foregin to host cell -DNA is more stable than RNA (spontaneous hydrolysis of RNA) -Mutation rate is higher in RNA, because RNA sequence repair mechanisms are less prevalent

what do dark reactions require?

-Requires reducing power, so the cell also needs to oxidize an external substrate (electron donor) to create NADH -Donor is not successively oxidized like in chemotrophy, but right at the membrane -Oxygenic photosynthesis: oxidation of H2O produces O2(cyanobacteria) -Anoxygenic photosynthesis: all other phototrophic bacteria (use H2S, H2,...)

Ethanolic Femrenetaiton

-Saccharomyces cerevisiae ("common baker's yeast) --Microbial eukaryotes --Single celled fungi; also called yeasts --Used in bread baking (sourdough) and alcoholic beverage industry --End products: CO2 and ethanol

Locations for nitrate (NO3-) = e- acceptor

-Terrestrial -Aquatic -Oxic and anoxic layers -Used up in upper sediment and soil layers just below the oxic zone

Calculating distance matrix form alignment

-The first step in making a tree is to align sequences -Second step is a distance matrix is calculated from the number of sequence differences -Third step is the tree is constructed by adding nodes to join lineages that have the fewest differences

human virome

-Their function as transduction agents may have major impact on bacterial evolution -May confer new metabolic or other beneficial properties (retained when beneficial in habitat)

Goals of systems biology

-Understand the functions of a specific organism on all possible molecular levels, and, ideally at all levels in its natural environment -Understand the structure and function of microbial communities on all possible molecular levels and, ideally, at all levels in their natural environments

Why did the nucleus develop?

-formation may be associated with evolution of RNA processing -Spliceosomes remove introns from eukaryotic mRNAs

Genotypic Analysis

-genome -Challenge: getting clean and complete genome information

phenotypic analysis

-morphological, metabolic, physiological, chemical characteristics -Challenge: requires the ability to culture, AND getting a pure culture

Genetic engineering include laboratory techniques such as

-nucleic acid hybridization. -molecular cloning. -restriction enzyme digestion.

Microarrays allow for

-the analysis of global gene expression. -the hybridization of cDNA (converted from mRNA) with DNA affixed to a solid-state support. -a comparison of expression of different genes under different growth conditions.

phylogenetic analysis

-using a gene marker showing evolutionary distance -challenge for unknown organisms PCR primers/probes may not bind

Heterofermentative

-yields lactate, ethanol, CO2 and 1 ATP/glucose --Lactobacillus brevis --Sauerkraut and pickles

Homofermentative

-yields lactic acid only and 2 --ATP/glucose --Lactobacillus delbrueckii --Milk and dairy products

What do light reactions produce?

ATP

Photoautotrophy requires

ATP production and CO2 reduction

What the organisms oxidize...

Chemoorganotrophs: are at top of pH scale Chemolithotrophs: are in middle of pH scale Phototrophs: bottom of the pH scale

Bacteria are mainly infected by

Class I viruses

Eukarya are mainly infected by

Class I, II, and IV

CRISPR

Clustered Regularly Interspaced Short Palindromic Repeats that protect bacteria from bacteriophage infection

__________ is a process that involves cell-to-cell contact to move the plasmid from a donor cell to a recipient cell.

Conjugation

Photosynthesis

Conversion of light energy from the sun into chemical energy.

Bacteria are mainly infected by _____ viruses, while animal cells are infected by _________ viruses.

DNA / RNA

Restriction enzymes recognize specific nucleotide sequences and cut them as part of a defense mechanism in bacteria. They are cutting

DNA double-strands

specialized transduction

DNA form a specific region of the host genome(genes), integrated among the virus genome

generalized transduction

DNA from any region of the host genome is packaged typically filling entire virion, random, often not whole genes

In order to apply PCR for DNA-based diagnostics, we make use of the principle that (a) _________ operates only when the last nucleotide of the __________ on the 3'-end can bind, so polymorphisms in a DNA sequence can be detected via this mechanism.

DNA polymerase / primer

Pandoravirus is the largest known virus, and has a _______ genome that is ________ than the smallest bacterial genome.

DNA/larger

Place organism in hierarchical system:

Domain Phylum Class Order Family Genus Species

Oxygenic photosynthesis

Employs 2 reaction centers, quinone-pool and Fe-S complex

Natural sources for mutations can be a result of

Error during DNA replication -most often, a single or few nucleotide is/are different

Which of the following statements is true?

Eukaryotic genomes are composed mostly of noncoding DNA, whereas bacterial genomes have mostly coding DNA.

A silent mutation is the most common single nucleotide change.

False

If a change in nucleotide sequence information does not change the cell's phenotype compared to wild type it is not considered a mutation.

False

Mutation rates of DNA-containing cells are higher than DNA- or RNA-containing viruses.

False

The "tree of life" that shows the three domains Bacteria, Archaea, and Eukarya is the phylogenetic reconstruction of the evolutionary relationship between all organisms, and thanks to modern genetic technology it is now considered 100% correct.

False

The reason for the variability of the influenza virus - resulting in the necessity of developing a new vaccine basically every replicative season (~12 months) - lies in the virus alternating between an enveloped and unenveloped virion.

False

With the onset of the human microbiome project we learned that our body does not contain many microorganisms, and that especially our intestinal tract has a very low density of bacteria.

False

What is a metagenome?

Genome sequences from all the organisms in a particular environment.

Characterizing large pools of molecules

Genomics Transcriptomics Proteomics Metabolomics Secretomics Interactomics

Goal of genetic engineering

Heterologous expression- expression of a gene in a different host

naturally competent bacteria

Streptococcus Bacillus Vibrio

Which of the following is NOT a palindrome (recognition site for a restriction enzyme)?

TAGGAT

Transduction

Transfer of DNA from one cell to another by a bacteriophage

A suicide vector encodes a gene that kills the host when expressed or selected for its function. Only vectors that have been successfully inserted with the gene of interest (suicide gene is disrupted) can grow under selective conditions.

True

Antigenic shifts in the influenza genome generates hybrid viruses with strongly modified surface proteins, triggering new outbreaks of influenza after long periods of apparent immunity.

True

Bacteria of distantly related lineages can have some highly identical genes because they have means to share part of their genetic information via horizontal gene transfer.

True

In transduction, DNA is accidentally moved from one bacterium to another by a bacteriophage.

True

One important advantage of eukaryotic cells as hosts for cloning vectors is that they already possess the complex RNA and posttranslational processing systems required for the production of eukaryotic proteins.

True

Restriction endonucleases are naturally occurring enzymes in bacteria to cleave foreign/viral DNA.

True

The core genome is the collection of genes that is shared by all sub-lineages of a group of interest, while the pan genome includes all variants in that sub-lineage, and also includes the genetic information of plasmids.

True

The lactate end product from lactic acid fermentation is only partially oxidized and can be used by other bacteria in syntrophy, e.g., Propionibacterium, which further oxidizes lactate into acetate, propionate and CO2.

True

The multiple cloning site of a vector is within the lacZ gene. During the ligation of the gene of interest into this site the lacZ gene gets disrupted. Addition of X-Gal allows for screening. What is a correct description of a color result?

White colonies means that the lacZ gene has been disrupted.

Phages are viruses that infect

bacteria

Assembly

bringing the fragments together like a puzzle (overlapping regions, coverage), forming scaffolds, and ultimately the entire genetic molecule

Which molecule is used for the hybridization to DNA that is bound on a microchip?

cDNA

CO2 is a potent greenhouse gas, and it

can be accessed as carbon source by many bacteria, including phototrophic bacteria, as well as phototrophic eukaryotes (algae and plants).

Which of the following is NOT a feature of nitrogen fixation?

can be performed by phototrophic bacteria only

Facultative aerobes

can use with oxygen or nitrate

Bacteria usually do not...

carry much "unnecessary" DNA ("genetic streamlining")

Taxonomy

characterization, classifying.ranking, naming organisms

pan genome

core + individual variants in a given sub-lineage, can also include (multiple) plasmids

Consider a mutation in which the change is from UAC to UAU. Both codons specify the amino acid tyrosine. Which type of mutation is this?

silent mutation

ssRNA

single stranded RNA complementary to mRNA

Some RNA viruses carry a ___________ RNA molecule that serves directly as mRNA - one reason why the infectious cycle of these viruses is initiated very __________.

single-stranded / rapidly

Agarose gel electrophoresis separates DNA fragments from each other based on

size

EcoRI is a restriction enzyme that recognizes a six base-pair sequence and creates ends.

sticky

Transcriptomics

study of RNA patterns -Total RNA extracted during a certain growth condition -Reverse transcription mRNA→cDNA (complementary strand) --DNA is more stable than RNA, easier to work with -Sequencing of all transcribed mRNA sequences -Map to genome to see which genes they are

Mitochondria are

the cell's respiration machinery (with O2)

mitochondria are:

the cell's respiration machinery (with O2)

lactic acid fermentation

the chemical breakdown of carbohydrates that produces lactic acid as the main end product

Which gene is used for the reconstruction of the phylogenetic relatedness of all living cells?

the gene for the RNA in the small ribosomal subunit, 16S/18S rRNA

Denaturation of DNA refers to the process when

the hydrogen bonds between double-stranded DNA weaken, so the DNA separates into the single strands.

Via genetic engineering, the vaccinia virus was altered to produce a foreign protein to be used as a vaccine against

the smallpox virus

Metagenomics

the study of genetic material recovered directly from environmental samples -looking at all DNA from an environment --Resolve metabolic capabilities --unraveling different identities

Restriction endonucleases cut specific sequence regions in DNA molecules. Which molecule(s) do you have to cut with the same restriction enzyme for the purpose of molecular cloning?

the vector and the insert

Endosymbiotic or parasitic microorganisms usually have very small genomes because

they streamline their genetic content, keeping only genes necessary to produce what is not provided by the host.

The process of importing free DNA into bacterial cells is known as __________

transformation

Which of the following methods may introduce foreign DNA into a bacterial recipient?

transformation, transduction, and conjugation

The bacteriophage Mu always integrates its entire DNA chromosome into a specific site (5-nucleotide sequence) of the bacterial chromosome, thereby creating direct repeats on either side of the inserted viral DNA. This process is called

transposition

Bioinformatics

use of computer and specially designed programs to store and analyze sequences and structures of nucleic acids and proteins

genetic engineering

using in vitro techniques to alter genes in the lab

Viral Evolution

viruses contain genetic material but are not cellular in nature and cannot reproduce. It is theorized that viruses are a precursor to life and "bridge the gap" between non life and the UCA

Two step oxidation across two types of bacteria, occurring in aggregates:

1. Ammonium-oxidation to nitrate Nitrosomonas 2. Nitrite-oxidation to nitrate Nitrospira

Identifying an ORF

1. Computer finds possible start codons 2. Computer finds possible stop codons 3. Computer counts codons between start and stop 4. Computer finds possible RBS 5. Computer calculates codon bias in ORF 6. Computer decides if ORF is likely to be genuine 7. List of probable ORFs

Steps towards a whole genome

1. Genome extraction, fragmentation and sequencing 2.assembly 3.annotation

combining the gene of interest and vector

1.Cut DNA with restriction enzyme 2.Add vector cut with same restriction enzyme 3.Add DNA ligase to form recombinant molecules 4.Introduce recombinant vector into a host

Anoxygenic Photosynthesis

1.Light generates high potential electrons from bacteria/chlorophyll in reaction center 2. Elections flow through the membrane, analog to respiratory membrane, and generate proton motive force

Genome extraction, fragmentation and sequencing

several methods, differ in price, fragment length, and accuracy (error rate)

Mycobiome:

60+ fungi present on skin, in oral cavity, and other moist surfaces

The Baltimore Scheme

7 classes taxonomy based on nucleic acids

missense mutation

A base-pair substitution that results in a codon that codes for a different amino acid. -leads tp replacement of amino acids in a polypeptide (eg. UAC to AAC).

Mutation

A change in a gene or chromosome.

silent mutation

A mutation that changes a single nucleotide, but does not change the amino acid created. -do not affect sequence of polypeptide or phenotype, are typically at third base of codon

nonsense mutation

A mutation that changes an amino acid codon to one of the three stop codons, resulting in a shorter and usually nonfunctional protein.

polymerase chain reaction (PCR)

A technique for amplifying DNA in vitro by incubating with special primers, DNA polymerase molecules, and nucleotides.

Conjugation

A temporary union of two organisms for the purpose of DNA transfer.

What is a suicide vector in molecular cloning?

A vector containing a gene that, when being expressed or selected for, kills the host. Only vectors that have been successfully inserted with the gene of interest can grow under selective conditions.

Which statement is true?

All organisms differ in their genome size, and, their amount of genes.

Human Genome Project

An international collaborative effort to map and sequence the DNA of the entire human genome.

What the organisms reduce... what they respire!

Anaerobic respiration Aerobic respiration

Host-virus defense mechanisms

Bacteria produce restriction endonucleases that cleave foreign DNA signature sequences

where are pigments located?

Bacteria/Chlorophyll pigments and accessory pigments are located within special membranes

mixed solvent fermentation

From cheap sources/waster Acetone, butanol, ethanol (solvent) production Microbial production of biofuels

chromosomal islands

Region of bacterial chromosome of foreign origin that contains clustered genes for some extra property such as virulence or symbiosis -Can be identified by genome comparison of close relatives --Clusters of genes for specialized functions not essential for survival --But can give selective advantage e.g., for biodegradation, nitrogen fixation, magnetosomes

gene transfer

Movement of genetic information between organisms

gel electrophoresis

Procedure used to separate and analyze DNA fragments by placing a mixture of DNA fragments at one end of a porous gel and applying an electrical voltage to the gel

nitrogen fixation

Process of converting nitrogen gas into ammonia

propionic acid fermentation

Propionibacterium and related prokaryotes produce propionic acid as a major fermentation product

There is scientific evidence that the first cell-like "virocells" and the first viruses on Earth contained _____ genomes.

RNA

What is NOT an ingredient of a PCR reaction?

RNA polymerase

Mutation rates

RNA viruses: 10^-3 DNA viruses: 10^-7 Bacteria: 10^-8 Eukarya: 10^-9

light reactions

The first of two major stages in photosynthesis (preceding the Calvin cycle). These reactions, which occur on the thylakoid membranes of the chloroplast or on membranes of certain prokaryotes, convert solar energy to the chemical energy of ATP and NADPH, releasing oxygen in the process.

What assumption is made for using a specific gene for the reconstruction of an evolutionary model of species divergence?

The gene has to be present in all species, and the mutation rate of the gene is constant.

Which of the following is NOT true of plasmids?

They are found in many eukaryotes.

The light reaction involves photosystems with light-active molecules, such as chlorophyll, that are localized ___________ of the photorophic cell.

in specially folded membranes

Point mutations

a single nucleotide is exchanged for another

Polyvalent vaccine

a single vaccine that immunizes against two different infections, e.g., an attenuated virus is also carrying an additional gene from another pathogen (attenuated + vector)

Phototrophic bacteria have a large variety of light-harvesting and accessory pigments, so they can _______ certain wavelengths of the visible light spectrum for energy generation.

absorb

nomenclature

agreed words and word stems to be used for the ranks

During genome assembly, you first have to __________, and then ________ .

align overlapping DNA fragments / ideally reconstruct the entire chromosome

Part of the development of modern-day Eukarya was that a cell of the archaeal lineage underwent endosymbiosis with a(n) _______, and turned it into a ________.

alphaproteobacterium / mitochondrium

In anaerobic soils

ammonium is respired (reduced) back to NH3 or to N2 gas -Ammonium is preferred nitrogen source for bacteria -Potential loss of N

In well-aerated soils

ammonium is turned over into nitrate by microorganisms -Nitrate is the preferred nitrogen source for plants -biofertilizer

Carboxysomes

inclusions with protein shell containing and improving efficiency of RubisCO in many autotrophs

Mutations happen randomly in the chromosomes of all organisms, and those that ________ the fitness of the organism in a given environment will be retained in the lineage.

increase

Genomics

discipline of sequencing,analyzing and comparing genomes

dsRNA

double stranded RNA

Most plasmids are

double-stranded DNA, though a few are not.

Cyclic photophosphorylation:

electrons move within closed loop; no net input or consumption

Larger genomes...

encode more transcriptional regulation and signal transduction

Smaller genomes...

encode more translational processes

RNA synthase

enzyme that catalyzes transcription process.

mutation rate

establishment of differences in nucleotide sequence in a population(diversification)

Annotation

find open reading frames (OFs)= "genes"; DNA sequence that codes for a polypeptide). Check nucleotide and amino acid sequence against databases, similarities suggest function of the ORF

how did chloroplasts arise?

from stable incorporation (endosymbiosis) of a cyanobacterium-like cell into the cytoplasm of a eukaryotic cell, leading to eukaryotic photosynthesis (algar, plants)

how did mitochondria arise?

from stable incorporation (endosymbiosis) of an aerobic respiring Alpha Proteobacterium-like cell into the cytoplasm of early eukaryotic cells

Vector vaccine

genes from a pathogenic virus is engineered to the genome of a harmless carrier virus, induces immunity to pathogen

core genome

genes shared by all sub-lineages

Lactate, ethanol and CO2 are end products:

heterofermentative/heterolactic fermentation

The study of gene origin can be extremely difficult in Bacteria, because they do not only pass genes to their progeny cells. Bacteria also perform a extensive amount of active

horizontal gene transfer.

Only lactate is end product:

hormofemrnetation/homolatic fermentation

Photosynthesis originated:

in bacteria -First: anoxygenic photosynthesis (not producing O2) -Later: oxygenic photosynthesis (O2 is a waste product)

molecular cloning

isolation and incorporation of a piece of DNA into a vector so it can be replicated and manipulated

DNA instead of thymine:

letal!

Systematics

links phylogeny (relationship of genes) with taxonomy (ranking this diversity of organisms with name categories)

attenuated vaccine

modified pathogen via genetic engineering, virulence factors have been deleted, but retained features to elicit immune responses (recombinant, infective)

Bottleneck

molecular biologists today need to speak computer language(s) and/or be able to work on the computer console (python, R, mothur,...)

Estimations state that the Earth's virome counts ________ virus particles than bacterial cells, and most of those viruses are _______.

more / bacteriophages

where does photosynthesis occur in bacteria?

pigments are integrated into complex invaginations of cytoplasmic membrane: --Vesicular chromatophores or lamellae in purple bacteria --Chlorosomes in green sulfur bacteria --Thylakoids in cyanobacteria

Proofreading activity is intrinsic to the __________ enzyme to _________ the accumulation of mutations during a replicative cycle.

polymerase / minimize

Herpesviruses are a large collection of different viruses that infect humans. Once infected, they typically remain latent in the body for an entire lifetime, and may break out occasionally (enter lytic stage). Where do these viruses reside?

nerve cells

New traits can evolve quickly in bacteria due to their high mutation rate and short generation time. But only mutations that are _______ or _______ for the fitness of the organism at a given selective pressure result in a retaining of the mutation(s) in the lineage.

neutral / favorable

Mutations cause

new genes and alleles to arise -heritable changes

Phototrophs

obtain energy from light -use energy from light to reduce CO2 to organic compounds

Deletions

one or more nucleotides are deleted

insertions

one or more nucleotides are inserted

During genome annotation, you first have to identify __________, and then ________ by comparison to databases.

open reading frames / assign functions to them

The genes individual to each strain within a bacterial species is called the

pan genome

The polyphasic approach to ultimately describe a true new species of Bacteria comprises a _________, _________, and _________ characterization.

phenotypic / genotypic / phylogenetic

where does photosynthesis occur in Eukarya

photosynthesis occurs in chloroplasts (intracellular organelles containing thylakoids: stacked, sheet like membrane systems)

A polymerase chain reaction (PCR) amplifies a certain genetic segment bordered on each end with a(n) ________

primer

autotrophy

process by which CO2 is reduced and assimilated into cells

Transformation

process in which one strain of bacteria is changed by a gene or genes from another strain of bacteria -Genetic transfer process by which free DNA is incorporated into a recipient cell and brings about genetic change

Sequencing of the mitochondrial genome indicates that mitochondria are most closely related to

proteobacteria

Purple sulfur bacteria

quinone pool as intermediate e-acceptor and H+-translocation

A molecule of the group of __________ is used as a "molecular clock" to determine the approximate time since two lineages of organisms diverged.

rRNAs

Carl Woese chose to examine phylogeny using rRNA sequences because

rRNAs are part of ribosomes - molecules present in all and function similar in all living organisms.

Calvin Cycle

reactions of photosynthesis in which energy from ATP and NADPH is used to build high-energy compounds such as sugars

The CRISPR system

recognizes foreign DNA sequences that have previously entered the cell and directs the Cas proteins to destroy them.

In the experiment of bacterial evolution we selected for the gain of function (ability to produce tryptophan) in those bacteria that have

recombined externally provided DNA into their chromosome.

dark reactions

reduces atmospheric CO2 to cell material for growth

The DNA of herpesviruses replicates by the __________ method.

rolling-circle


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