Cell Division

Dewey Nguyen

Mitosis – somatic/body or boring cell division!
• Two identical daughter cells, just like parent cell • Diploid, 2n, two copies of each chromosome

Interphase- Prep phase for mitosis
• G1 – make organelles and produce macromolecules

• S –DNA is decondensed form of chromatin,DNA replication
• G2 – additional growth

The 5 Mitotic Phases .Prophase • Formation of mitotic spindles • Centrosomes are moving to opposite ends • Sister chromatids are condensed .

Prometaphase • Nuclear envelope is broken down • Kinetochore microtubules are attached to centromeres • Non-attaching kinetochore microtubules elongate the cell .

kinesin e) Actin. dynein b) Postive.Question The ____ end of the kinetochore microtubules requires _______ to pull the chromosomes toward the plasma membrane. dynein d) Negative. kinesin c) Negative. myosin . a) Positive.

nuclear envelope formation .The final steps • Metaphase plate formed in metaphase • Anaphase – sister chromatids are separated • Telophase and Cytokinesis – cleavage using actin filaments and myosin.

Question Sister chromatids would first be seen at this stage: a) G1 b) S c) Prophase d) Metaphase e) B and C .

how would this drug affect mitosis? a) The cell cannot elongate for cell division b) Sister chromatids cannot be separated c) Cytokinesis cannot take place d) A and B e) None of the above .Question A drug is found to disrupt the function of kinesin.

Oh Meiosis . one copy of each chromosome • All stages are very similar to Mitosis .the sex cell division • Four different daughter cells • Haploid. n.

Oh Meiosis • Metaphase I – homologous chromosomes formed tetrads and crossing over occurs here! • Independent assortment.Metaphase I and II – increases genetics variation .

Sister… homologous … what!?!?

Oh … I get it!
• Sister chromatids are the exact same thing! • Homologous chromosomes carried the different variations of the same gene! • Non-homologous chromosomes are different chromosomes!

Crossing-Over … is what?!?!
• Yes, crossing-over is chromosomal sex, where genetic materials are exchanged! So sister chromatids cannot do it together! Only do it with the same chromosome or homologous chromosome! • Must hold hand first, so tetrad are formed, and then … Chiasmata! Let’s take a closer look!

So this is how they do it! .

To create more variations! . each alleles separated into gametes independently Wait… what is the point of these two law? .Mendel • Law of segregation into two different gametes • Law of independent assortment – Two alleles will separate – When two or more characters are inherited.

Test Cross • To test if a phenotype is homozygous dominant or heterozygous dominant. • Cross with a homozygous recessive .

Non-Mendelian Inheritances • Incomplete dominance • Co-dominance • Multiple Alleles • • • • Pleiotropy Epistasis Polygenic inheritance Environmental factors/impacts .

Non-Mendelian Inheritances • Let’s say R = red. and r = white. So Rr would yield: – Incomplete dominance – Co-dominance .

Recombination Map units … don’t add up! Offspring show different phenotype from parents! .

Abnormalities • Karyotopic Abnormalities – having an abnormal number of chromosomes • Chromosomal aberrations – a part of a chromosome is messed up! – Only translocation can involve two chromosomes .

and your sister is also color blind. but you are not.Question Your father is color blind. What is the probability that you will have a diseased brother? a) 0 b) 1/2 c) 1/4 d) 1/8 e) None of the above .

and your sister also has achondroplasia. but you are not. What is the probability that you will have a diseased brother? a) 0 b) 1/2 c) 1/4 d) 1/8 e) None of the above .Question Your father has achondroplasia.

Question A female duck has a mosaic feathers. what could be one explanation for this? a) Barr body b) Incomplete dominance c) Co-dominance d) Epistasis e) More than one possible answer .

Bio Sci Peer Tutoring FINAL Review: Bio93 Sec A+B Arun Manmadhan UNIT VI: Molecular Biology .

Frederick Griffith .

So what did this show/mean?? • S (Smooth) cell = DEADLY ▫ smooth slimy coat gives the bacteria protection • R (Rough) cell = Harmless • Heat killed S cell = Harmless ▫ The cell is killed but not the DNA • Heat killed S cell + R cell = DEADLY ▫ DNA from heat killed S cell „transforms‟ the R cell into an S cell • KEY POINT: Showed that there exists a „transforming principle‟ but not necessarily DNA. RNA or Protein .

Alfred Hershey and Martha Chase Experiment
• Showed that DNA was the transforming agent and thus the unit of hereditary.
• Phage = a virus • SULFUR is FOUND in Protein but NOT in DNA • Phosphorous is FOUND in DNA but NOT in Protein

HELICAL structure . Watson & Crick • Rosalind Franklin used X-ray crystallography to get a real picture of DNA • Watson and Crick then analyzed the picture to come up with a structure that FIT the data • Basic Postulates: 1. DNA base pairing rules A=T.Rosalind Franklin. DNA is LINEAR 2. C ≡ G 3.

Properties of DNA • Purines = Guanine (G) and Adenine (A) • Pyrimadines = Thymine (T) and Cytosine (C) • Phophate group = 5‟ end • Sugar group = 3‟ end • DNA is anti-parallel one strand = 5‟3‟ the other = 3‟5‟ • DNA replication is SEMI-CONSERVATIVE • DNA is replicated with DNA POLYMERASE .

the second strand (red) is new Semiconservative model: Type of DNA replication in which the replicated double helix consists of one old strand. derived from the old molecule. and one newly made strand .Definition G-33 One parental is Preserved.

Question In the Hershey and Chase experiment with the bacteriophages. the most important clue to the chemical nature of the hereditary material was the: a)incorporation of radioactive sulfur in the bacterial genome b)incorporation of radioactive phosphorus in the bacterial genome c)transformation of R cells to S cells via heat killed S cells d) accumulation of phosphorus on the surface of the bacteria .

Question If one strand of a DNA molecule has the base sequence 5‟-ATTGCAT-3‟. its complementary strand will have the sequence: A) B) C) D) E) 5‟-ATTGCAT-3‟ 5‟-ATGCAAT-3‟ 5‟-TAACGTA-3‟ 5‟-CGGTACG-3 None of the above .

Solution • Sequence given: ▫ 5‟-ATTGCAT-3‟ ▫ 3‟-TAACGTA-5‟ This is the complementary strand But since all options are written 5‟3‟ flip it! ▫ 5‟-ATGCAAT-3‟ Same as choice B .

The Central Dogma DNA • DNA RNA= Transcription mRNA • 5‟ cap • Poly A Tail • RNA Splicing Protein • RNA  Protein = translation Lecture 22 .

Prokaryotes vs Eukaryotes Eukaryotes Prokaryotes MORE COMPEX! .

Transcription • Initiation: Promoter region + transcription factors • Promoter region = DNA sequences which provide a binding site for RNA polymerase • Transcription Factors= recruit RNA pol to the promoter region .

Transcription continued • Elongation = RNA polymerase adds RNA nucleotides until it hits a termination „spot‟ • Termination = RNA polymerase disassociates from the template DNA strand and primary transcript is released to be processed. .

Question In order for a particular drug to directly inhibit RNA polymerase activity in animals it must have: A) Non-polar functional groups B) Nuclear import signal C) Compatible transcription factors D) Complementary sequences to the promoter E) None of the above .

Question Promoters for eukaryotic mRNA synthesis: A) are more complex than prokaryotic promoters B) can require binding of multiple transcription factors to form a transcription complex C) have specific DNA sequences such as the "TATA" box that are recognized by proteins D) are the stretches of DNA to which RNA polymerase binds to initiate transcription E) All of the above .

Everything occurs in the cytoplasm Transcription + RNA processing in the NUCLEUS Translation in CYTOPLASM Bacteria! YOU & ME!! .

the mRNA is enzymatically degraded.important for the nuclear export.RNA processing • 5‟ cap added . ▫ Adenosine (A) repeats .Capping ensures the messenger RNA's stability while it undergoes translation ▫ Mostly Guanine (G) repeats • Poly A Tail . The tail is shortened over time and when it is short enough. translation and stability of mRNA.

RNA splicing exon intron Exon INTrons = INTerfere Primary Transcript Intron Exon Intron + 5‟cap + poly A Tail Introns REMOVED “Mature” Transcript 5‟ cap Exon Exon Exon Poly A Tail .

UGA. • The code is nearly universal. and only one amino acid.Genetic Code • Memorize!!!: ▫ Start Codon: AUG ▫ Stop Codons: UAA. • The genetic code is degenerate. UAG • The genetic code is read in triplets (codons) • It is non-overlapping • The genetic code is unambiguous ▫ Each codon specifies a particular amino acid. ▫ each amino acid can be specified by more than one codon. .

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Question Which of the following is NOT a feature of eukaryotic gene expression? A) Random repeat segments at mRNA poles B) many genes are interrupted by non-gene coding DNA sequences C) RNA synthesis and protein synthesis are coupled D) mRNA is often extensively modified before translation E) Addition of a nuclear export tag to mature mRNA .

tRNA has the complementary anti-codon: 3‟-UAC-5‟ • Ribosomes read mRNA in 5‟3‟ direction • E site: Exit site. tRNA‟s leave • P site: Peptidyl site.Translation – important points • tRNA molcules are the translators • Purpose??? MAKE PROTEIN. AND MORE PROTEIN!!!!! • Posses ANTI-CODONS to the CODONS found in mRNA • Thus if mRNA has codon: 5‟-AUG-3‟. amino acid chain lives/ grows here • A site: Acceptor. temporary home for tRNA‟s . PROTEIN.

What does that spell?! • E-P-A • Environmental Protection Agency .

Mutations – Oh Nooooo! • Base pair substitution Normal “Wildtype” 5‟-AUGAAGUUUGGCUAA-‟3 Mutant 5‟-AUGAAGUUUGGUUAA-‟3 • Types: ▫ Silent: mutation has no effect on amino acid added b/c genetic code is DEGENERATE ▫ Missense: mutation changes one amino acid because of mutation ▫ Nonsense: mutation results in a STOP codon. prematurely cuts the amino acid chain .

the mutation is a FRAMESHIFT • Wildtype: ▫ THE|FAT|CAT|ATE|THE|WEE|RAT Insert ONE or TWO Nucleotides • Mutant (frameshift mutation): ▫ THE|FAT|CAT|ATE|THZ|ZWE|ERA|T .Mutations continued… • If mutations add or remove nucleotides in anything BUT multiples of three.

Question A nonsense mutation generally results in: A) B) C) D) Translation of incorrect amino acids a shorter polypeptide no change in expressed protein a frameshift .

Question Which of the following is NOT correct? A) There are sixty-four different codons. B) All codons specify a specific amino acid. C) Some codons are used for initiation or termination D) There are more codons than amino acids E) All of the above are correct .

DNA Cloning In Bacteria First Step: Isolate DNA/Gene of interest Second Step: Insert DNA into a PLASMID Third Step: Allow Bacteria to incorporate DNA and multiply Fourth Step: Observe/ Extract protein of interest synthesized .

Cutting DNA • You need palindrome sequences IE racecar ▫ Read from both directions = same word • Restriction enzymes cut DNA at these palindrome sequences • Create Sticky Ends • Once DNA has been inserted into a plasmid. covalent bonds are reformed using DNA Ligase .

.PCR Reaction • Start with one piece of DNA • Can amplify and duplicate millions and billions of copies in a few hours.

Allows for identification of certain Sequences of DNA.Gel electrophoresis Purpose: Separate DNA sequences Based on size. DNA is negatively charged. What makes DNA NEGATIVELY Charged???? . so moves To positive end. Smaller DNA Molecules feel less resistance in the Gel and thus move a greater distance.

Protein modification Protein stability/degradation. RNA translation. 4. . RNA stability/degradation. RNA processing. 2.Continued (Copied from your lecture notes) KNOW THIS! 1. Gene expression can also be regulated at many other steps such as 1. RNA transport. 6. 5. A SMALL fraction of the genes in the genome are expressed in any given cell 2. 3.

• Micro RNA (miRNA) – attach to MATURE mRNA and either DEGRADE or PREVENT TRANSLATION .Continued • Transcription Factors: Tell the cell/ polymerases WHEN and WHERE transcription can occur • Enhancers: Work with transcription factors and the promoter region to stimulate transcription.

Gene Expression So MANY check points that can control how the gene is expressed .

C) DNA polymerase .production of DNA fragments for gene cloning B) DNA ligase .enzyme that cuts DNA.detect if bacteria transformed . creating sticky ends.copies DNA sequences in the polymerase chain reaction D) Electrophoresis .Question Which of the following tools of recombinant DNA technology is INCORRECTLY paired with one of its uses? A) restriction enzyme .Detect DNA length E) Antibiotic resistance .

Question Which of the following is not part of the normal process of cloning recombinant DNA in bacteria? A) restriction enzyme digestion of cellular and plasmid DNAs B) production of recombinant DNA using DNA ligases and a mixture of digested cellular and plasmid DNAs C) separation of recombinant DNAs by electrophoresis D) transformation of bacteria by the recombinant DNA plasmids E) All of the above are valid processes .

Time permitting questions… The regions of DNA in a eukaryotic gene that encode a polypeptide product are called: A) promoters B) Exons C) Introns D) tRNA's E) Transcription factors .

serving as a template for translating. respectively are: A)mRNA.mRNA E)rRNA. and transporting amino acid.mRNA.rRNA D)tRNA.tRNA.rRNA B)rRNA.mRNA C)tRNA.tRNA.tRNA .mRNA.Question Three types of RNA involved in comprising the structural and functional core for protein synthesis.rRNA.

Question A messenger acid is 336 nucleotides long. The number of amino acids in the protein translated from this mRNA is: A) 999 B) 630 C) 330 D) 111 E) 112 . including the initiator and termination codons.

Question A key feature for the identification of plasmids containing recombinant DNA is *hint white vs blue colonies): A) weighing it B) the DNA sequencing of recombinant plasmids C) the production of restriction maps of recombinant plasmids D) introns can be moved to new locations within the gene E) the disruption of a gene on the plasmid by the inserted recombinant DNA .

com/olc/dl/120076/bio21.html • http://bcs.htm • http://highered.com/webcontent/animations/content/meselson.com/sites/0072556781/student_view0/chapter14/animation_quiz_1.html • http://www-class.sumanasinc.html .swf • http://www.com/sites/0072556781/student_view0/chapter11/animation_quiz_4.html • http://highered.com/thelifewire/content/chp12/1202001.html • http://bcs.com/thelifewire/content/chp14/1402001.unl.edu/biochem/gp2/m_biology/animation/gene/gene_a3.mcgrawhill.html • http://highered.com/college/biology/animations/ch12a01.whfreeman.whfreeman.mcgrawhill.mcgraw-hill.Recommended Animations To Watch (copy and paste into browser each link separately) • http://nortonbooks.

Developmental Biology Kevin Huang M.edu . F 2-3 PM SH 149 kihuang@uci.

When zygote divides.D. My interpretation of CD's: essentially establish themselves during early divisions of cleavage (You likely don't have to know that bicoid is an example of C.: Common examples that you guys see cell induction could extend to the actions of organizers which secrete signals (morphogens) to influence differentiation! .'s but it doesn't hurt for me to tell you)  Cell induction: the result of intercellular communication.I. Signals from one cell influence the fate of another cell.Developmental Theory A combination of ―Cytoplasmic Determinants‖ and ―Cell Induction‖  Cytoplasmic Determinants: are present in egg (maternal inheritance) before fertilization. My interpretation of C. different determinants yield different cell lineages.

Positional Information  Pattern formation: the establishment of axes A/P.How does this make sense? Well. D/V. HENCE. you know bicoid is a C.D.Pattern formation vs. Morphogens (like bicoid) yield a concentration gradient.  “Dumbing it down”: positional information --> This definition likely doesn't “make sense” so think of it in terms of how morphogens work. and determines A/P polarity! Positional Information: directs pattern formation by giving positional values to cells. P/D (determined by maternal cytoplasmic determinants present in the egg before fertilization). Different levels of bicoid yield different cell types. DIFFERENT .

cytoplasmic determinants are present in the egg before fertilization.Drosophila and ―bicoid‖  In drosophila. The resulting activation initiates a bicoid gradient across the embryo (highest at anterior. Fertilization results activation of the egg. lowest at posterior) High [bicoid] = head Intermediate [bicoid] = thorax    Bicoid acts as a morphogen .

forms germ layers (ectoderm. mesoderm. each cell becomes smaller.Overview Fertilization  Fusion of haploid egg/sperm to form a diploid zygote (influx of Ca2+ results in egg activation) Rapid cell divisions without an increase in overall size. Cell movement. Thus with each successive cleavage division.Embryonic Development . endoderm) ―formation of organs‖ (formation of neural tube to become future spinal cord/brain) Cleavage  Gastrulation  Organogenesis  .

Fertilization haploid sperm + haploid egg = diploid zygote Entry of sperm results in an influx of Ca2+ into the egg. (acrosomal rxn occurs before cortical rxn) . Egg is activated at this point. etc. Acrosomal rxn: there are specific receptors for the sperm. This results in increased protein synthesis. This essentially prevents interspecies fertilization Cortical rxn: essentially prevents polyspermy by blocking the entry of additional sperm.

At the end of cleavage a blastula is created. - the blastula has a cavity within it called the blastocoel . Involves duplication of organelles and DNA but no overall growth!!! Form of mitosis (once again: NO GROWTH) - - Morula is formed during cleavage.Cleavage The initial divisions of the embryo Rapid cell divisions that proceed after fertilization.

and the archenteron is characteristic of gastrulation. .Gastrulation Formation of germ layers Dorsal lip invaginates into the interior of the embryo in a process called involution. The blastocoel shrinks and in its place another cavity called the archenteron is present Blastopore lip encircles the yolk plug Generates the 3 germ layers: Ectoderm (outer) - - - Mesoderm (middle) Endoderm (inner) Because not a lot of background information is given to you guys—Remember that the blastocoel is characteristic of the blastula.

spinal cord) How to remember this: You know that the neural plate is ectoderm. you see that endoderm is the primary cell tissue type within the gastrula! It essentially invaginates to become the gut) . not super specific!) • Ectoderm • • epithelial/outer covering like the skin (easy to remember) Central nervous system (brain. and receives signals from the notochord to become FUTURE spinal cord/brain!! • Mesoderm • Notochord (you know that the notochord is mesoderm and signals to the overlying ectoderm—neural plate) Muscular. circulatory • • Endoderm • Gastrointestinal tract (looking at diagram on previous slide. skeletal.Know the 3 germ layer fates (general.

endoderm) develop into rudiments of organs Vertebrate organogenesis: neural plate (ectoderm).Organogenesis ―creation of organs‖ • Germ layers (ectoderm. mesoderm. notochord (mesoderm) – Mesoderm surrounding notochord → somites (which split to become body cavity) Neural fold Neural plate • Neural crest Neural crest Outer layer of ectoderm Neural tube .

CNS Formation (Brain/Spinal Cord) Organogenesis • Notochord (mesoderm) signals to overlying neural plate (ectoderm) which results in the inward folding of the plate---forming the neural tube (future brain/spinal cord) • Requires microtubules. actin .

Somites Key to early embryonic segmentation • • Not a lot is told to you guys about somites What I think you should know: – – Somites = mesoderm surrounding notochord Shows segmentation Somites HEAD Neural tube Head mesoderm Neural folds Tail .

the organizer will cause surrounding cells to change their destiny • Ex: you graft a ―foot organizer‖ where hands normally form---you will get feet for hands! • • 2) apical ectodermal ridge (AER) – Limb bud pattern formation 3) Zone of Polarizing Activity (ZPA) . they enforce their demands‖ What do I mean by this? • Organizers to know: • 1) dorsal lip of blastopore – notochord/neural tube formation They secrete factors (morphogens) that affect gene expression in surrounding cells If you graft organizers of particular regions into foreign regions. Wherever they are present.limb bud pattern formation .Organizers Important stuff! • Organizers act sort of like ―command centers‖.

AER/ZPA • ZPA secretes Sonic hedgehog which gives positional information posterior to anterior. and two ring fingers :O) . AER is overlying ectoderm • • ENGRAFTMENT OF NEW ZPA YIELDS POSTERIOR IDENTITY AT NEW LOCATION (notice in pic. That there are two pinkies.

Organizers ―anatomic identity‖  What to know:  HOMEOTIC GENES: Code for ―anatomic identity‖ (ex: tells a particular segment ―you are going to be a hand‖) ORGANIZERS: secrete signals that influence gene expression in surrounding cells  Homeotic is at the GENE level Organizers at the CELLULAR level .Homeotic Genes vs.

. Zygote (result of fertilization) Morula (during cleavage divisions) Blastula (the result of cleavage divisions) Gastrula (result of gastrulation) Neurula (result of neuralation)----this is known in our lectures as our example of ORGANOGENESIS... ...therefore know that the neural tube folding is during organogenesis! Blastomeres are the individual cells that make up the morula and blastula during cleavage.Tidbits you might want to be aware of....

Discussion Generation: The questions I present here are intended to broaden your understanding and help see the ―bigger picture‖ if development and how they may apply to other topics in the class :) Do organizers function as cytoplasmic determinants or cell induction? What kind of signals to they secrete? ―harder‖ question: How do organizers affect homeotic gene expression? Be as specific as possible! Another ―harder‖ question: How do organizers change the expression of cells in the area in which an engraftment has occured? .

Although the acrosomal rxn functions to prevent interspecies fertilization. Hint: this does not relate towards development.) real example: a ―mule‖ (horse + donkey) A mule is formed by the mating of a horse and a donkey. If I told you the # of chromosomes in a horse was 64. Ex: the ―zorse‖ (buahaha. think cell cycle.. sometimes it does not work indefinitely. . and in a donkey 62. however the species were compatible enough that an embryo was viable enough to develop.creepy.. . This means in this situation acrosomal rxn function failed. and in a mule 63---why do you think mule are sterile (cannot mate)? Random and irrelevant fact: when I looked up these values it turned out to be these exact numbers. .

What could be the best way to define the sort of signaling that cytoplasmic determinants and morphogens convene? .

what kind of long term effects can this have on the organism? .What if you could inhibit all mesoderm signaling to the ectoderm? Based off of what you know.

.THE NERVOUS SYSTEM Wendy’s fun-filled introduction to Neurons Brains are cool! Girls love brains.

Cl-. K+.Label the Neuron Indicate where the concentration of the following ions are highest in the neuron: Na+. and anions- .

Label the Action Potential A: B: 1: 2: 3: 4: .

Will the ion move in or out of the cell using these channels: 1. Voltage-gated K+ Channels ____________ 3. Non-gated K+ Channels ______________ . Voltage-gated Na+ Channels ___________ 2.

Label the Synapse A: B: C: D: E: F: G: H: .

d. b. Dendrite Axon hillock Synaptic Terminal Axon Node of Ranvier . e.What part of the neuron relays signals from one neuron to another neuron? a. c.

A nerve A neurotransmitter An axon A dendrite White matter . a. e. c. d. b.The part of the neuron that carries an impulse towards the cell body is called ________.

Graded changes in polarization e. b. The difference in voltage between the inside of the cell and the outside of a cell c.The membrane potential refers to: a. Ion gradients in excitable cells d. The voltage inside a cell during an action potential. The fact that neuronal cells must generate more membrane when they grow .

increasing the permeability of the membrane to Na+ b. stimulating the sodium-potassium pump . allowing Cl– to enter the cell e. insulating the hillock region of the axon d.An EPSP (excitatory postsynaptic potential) facilitates depolarization of the postsynaptic membrane by: a. increasing the permeability of the membrane to K+ c.

determine if the neuron membrane will be hyperpolarized. Binding of serotonin to ligand gated K+ channels __________________ After summation of inhibitory postsynaptic potentials _______________ .For each statement. depolarized or no change.

What is the approximate threshold? _________ Which letter(s) represent inhibitory post-synaptic potential? ________ Which letter(s) represent hyperpolarization? _______ The cell is depolarizing at letter: _________ Letter A is an example of? _________ .

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