UNIT SIX:  DNA, THE HEREDITARY MATERIAL; PROTEIN SYNTHESIS

Upon completion of this unit you will be able to:

1.  List several characteristics which the genetic material must possess.
2.  List the components of a nucleotide.
3.  Name and describe the five different nitrogenous bases.
4.  Explain how DNA molecules vary in structure from one species to another; thus
     producing  variations in the proteins of different species.

5.  Describe the bonding between the nitrogenous bases in the DNA molecule.
6.  Diagram the process of DNA replication if given a "sense" strand of DNA.
7.  Provide the functions of endonuclease, exonuclease, polymerase and ligase.
8.  Identify the different types of RNA.
9.  Diagram the process of DNA transcription if given a "sense" strand of DNA.
10.  Diagram the process of translation (protein synthesis).
11.  Compare and contrast DNA and RNA.
12.  Describe how DNA controls the synthesis of proteins in a cell.
13.  Differentiate between frameshift and point mutations.
14.  List several causes of mutations and how they might effect DNA.
15.  Distinguish between somatic and sex cells.
16.  Explain how cancer is caused and treated.
17.  Relate the operon concept to the regulation of gene activity and the functioning
       of a living organism.

18.  Define and discuss differentiation.
19.  Discuss bacterial plasmids and processes involved in recombinant DNA procedures.
20.  List the importance of transduction and transformation in bacteria.
21.  Define locus, gene and chromosome.
22.  Describe the chromosome theory.
23.  Differentiate between haploid and diploid cells.

Unit References:  Text Chapter 3 pgs. 56-59; Text Chapter 8
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Unit 6:  DNA The Hereditary Material/ Protein Synthesis

DNA- DeoxyriboNucleic Acid;  reference library of "How-To" books

I  Search for Genetic Material
     A. Requirements for genetic material:   (Objective #1)

          1.  Must be able to store info used to control development and metabolic activities.

          2.   Must be stable so that it can be replicated w/ high fidelity and passed generation
                to generation
          3.   Must be able to undergo mutation to generate variability  (changes must
                be inheritable)
          4.   Must be chemically diverse

          5.   Must use stored info to direct synthesis of products necessary for operation of cell.

     B. Early researchers believed protein was genetic material because of its diversity.
          1. Meischer- nuclein

          2. Griffiths (1928)  bacterial transformation, material passed

          3. Avery and others- chemical proof that Griffiths found DNA not protein
          4. Hershey and Chase (1952)- used virus and radioactive isotopes of sulfur
              and phosphorous, Concluded:  DNA genetic material

          5. Watson/Crick/Franklin (1953)- structure of DNA, double helix w/ 4 nitrogenous bases
               of differing sequences.

II.  Structure of DNA (Objectives #2, 3)  Text pg. 56-59, 144-147  

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                                    What Is A Genome?

     A. DNA- deoxyribonucleic acid
          1. Double Helix (zipper, ladder, spiral staircase)
          2. Helix consists of 2 complementary chains of nucleotides
          3. Nucleotide = 1 nitrogenous base + 1 deoxyribose sugar + 1 phosphate

          4. Nitrogenous bases:     PURINES             PYRIMIDINES
                                                     Adenine (A)         Thymine (T)
                                                     Guanine (G)         Cytosine (C)

          DOUBLE STRANDED HELIX  (Objective #5) Text pg 145
                                                       Hydrogen bonds

                                                       Covalent bonds

                                                       S- deoxyribose sugar

                                                       P- phosphate
 
 
 
 

     B. RNA- RiboNucleic Acid  Text pg. 58, 146-147
          1. uses ribose sugar rather than deoxyribose
          2. single stranded
          3. Bases  A, C, G, and U (uracil, substitutes for thymine)

     C. Chargaff's Rules-  Complementary Base Pairing  (Objective #4, 5)
          1. Base composition of DNA differs from species to species

               a. amount of DNA
               b. sequences of bases on DNA

          2. In each species, however, the %A = %T and %C = %G

               -further, 50% of bases were purines (A & G) and 50% of bases were
                pyrimidines (T & C)
               -therefore, A must pair w/ T; C must pair w/G

     D. DNA Replication - mitosis (cell division)  (Objective #6) text pgs. 145-146
          -Watson/Crick model suggests  replication by  Complementary Base Pairing
         -during replication each parental DNA strand serves as a template ("blueprint")
            for a new strand
          -replication requires the following steps:

          1. DNA must unwind (helicase) and "unzip" (break hydrogen bonds)

          2. Complementary nucleotides, always present in nucleus, move into place by
                complementary base pairing

          3. Nucleotides joined to reestablish double strand:  Steps #2, #3 carried out by DNA
                Polymerase
          4. When done, 2 completed, identical, double stranded DNA molecules present
 
 
 
 
 
 
 
 
 

DNA Replication is Semiconservative- each new double helix contains one parental strand and
        one  new strand;  Meselson and Stahl 1958- radioactive isotope N 15
 
E.  Enzyme Activity  (Objective #7)
          1. Polymerase- enzyme responsible for addition of nitrogenous bases (nucleotides)
                to newly   forming strands of DNA
               a. Has proofreading function that checks each pairing.

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          2. Exonuclease- can remove nucleotide from end of chain
 
 
 

          3. Endonuclease- break bonds within chain (restriction enzymes)
 
 

          4. Ligase- repairs nicks and cuts within chain
 
 
 
 

F. Accuracy of Duplication
          -estimated mistakes 1/10,000 base pairs
          -actual 1/1,000,000,000
               much better than estimated due to action of polymerase proofreading
          -if error does occur and is not corrected, then a gene mutation has occurred.

II.  PROTEIN SYNTHESIS- (Objective #9, 10, 12)  Text pgs. 147-155
-gene expression, cell activity; DNA master code for protein synthesis

          Problem:  DNA in Nucleus --> protein synthesis occurs in cytoplasm, how do
                instructions get to construction site?

DNA --> copy small piece --> remove from nucleus --> take to ribosome --> let workers read
instructions and assemble --> protein (gene product)

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                                    Role of the Nucleus in Protein Synthesis
                                    Molecule of the Month - Ribosome
                                     DNA - RNA - Protein
                                    Transcribe and Translate a Gene

(Objective #8)
     A. RNA- RiboNucleic Acid; nucleotides like DNA except:
          1. Ribose rather than deoxyribose sugar
          2. Pyrimidines (cytosine and uracil) RNA uses U rather than T
         

     B. Part I:  Transcription, making of mRNA- messenger RNA (Text pg 147-148)
          -"Reads", copies or TRANSCRIBES (transcription) DNA code, takes message (copy)
                to cytoplasm

     C. rRNA- ribosomal RNA; Ribosomes
          -site of protein synthesis, RIBOSOMES made of RNA
          -site where code is translated (decoded)

     D. Part II Translation, using tRNA- transfer RNA to translate mRNA code
            3 parts:  Initiation, Elongation, Termination (text pgs 148-151)

          -clover leaf shape- string of paired and unpaired nucleotides
          -translates code contained on mRNA (Text 148-151)

               1. every 3 bases in DNA (or mRNA copy) codes for one Amino Acid

               2. tRNA translates and carries appropriate amino acid to ribosome

               3. at ribosome tRNA deposits amino acid coded for and A.A.chain (protein)
                    is created.

     E. THE PROCESS: DIAGRAM FORM (Text pgs 150-154)
 
 

DNA vs. RNA  (Objective #11)

                                  DNA                                                           RNA
FUNCTION            genes, controls protein                   helper to DNA, involved in
                                synthesis                                          actual building (synthesis) of protein

SUGAR                  deoxyribose                                     ribose

BASES                  A, C, T, G                                        A, C, U, G

STRUCTURE       double stranded;                             single stranded
                               complementary base
                               pairing; helix

                                                T    A    C    A    T    G    G    G    T    C    A    T    A    T    C

REPLICATE ---->

TRANSCRIBE --->

TRANSLATE ---->
 

MUTATIONS- (Objective #13, 14)  Text pgs.158-160 -  an inheritable change in a genetic
character resulting either from a change in a gene at a specific point or an alteration of
chromosome structure.

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     1. Point Mutation - a mutation that involves a single base pair.  Usually a substitution.

          -may result in no noticable change or code for different protein (amino acid)
            resulting in something as serious as PKU or Sickle cell

          -Spontaneous Mutations
               Tautomeric shift- a shift in the form of a nitrogenous base

     2. Frame Shift- removal or addition of a base pair, alterations effect everything after
            the addition or deletion
 

EX.      POINT MUTATION                              FRAME SHIFT MUTATION

          THE CAT ATE THE RED HAT             THE CAT ATE THE RED HAT

          THE RAT ATE THE RED HAT             THE ATA TET HER EDH AT (DEL C)

          THE CAT ATE THE RED CAT             THE CCA TAT ETH ERE DHA (ADD C

     3. Causes- mutagens- environmental substances that cause mutations (text pg 171-173)

          Radiation-     Ionizing and X-ray ---> Break DNA, causes deletions and  translocations

                         U.V. ---> alters DNA of surface cells (thymine dimers)

                         **cause primarily Frame Shift type mutations

          Organic Chemicals:  Pesticides
                              Cigarette smoke (benzo(a)pyrine) and 9 others
                              automobile exaust
                              Smoked foods
                              Agent orange/mustard gas - methylating/ alkylating groups added to DNA

                         **all cause mispairing of bases (Point Mutation) usually a single nucleotide
                            substitution

(Objective #15)
               -if mutation in somatic cells (body cells) ---> cancer
               -if mutation in gametes (sex cells) ---> genetic defect in offspring

CANCER (Objective #16)  Text pg 171-173   abnormal division of cells; when cells rebel!
     Carcinogens- noxious chemicals, radiation, virus

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          -cause alterations in DNA which cause cells to divide uncontrolably
            (environmental/ self imposed)

          -Oncogenes- stretches of DNA capable of transforming normal body cells into
            tumor cells.

          -p53, Apoptosis

          *cancer arises when genetic material damaged in a manner that frees the cell from
            normal constraints of growth

     TUMORS- clusters of cancer cells; some grow fast, some grow slow
          Benign ("kind")- growth restricted to area of tumor
          Malignant ("evil")- breaks from original mass, enabling them to metastasize
                (spread through  body)

          Treatment- center around removal of tumor (early detection key)
               1. Surgery- location dependent
               2. Chemotherapy- lowers immune reactions and others
               3. Radiation- effective on rapidly dividing cells, however, . . . . .
               4. Immune therapy- build bodies own defenses
 

BACTERIAL DNA  (Objective #19, 20)  -  Pioneering Modern Biotechnology
    
     A. Transformation- Free DNA in environment becomse incorporated in bacterial cells
            (Griffiths,  Avery)
          -genetic change in one strain of bacteria brought about by exposure to freshly killed
            bacteria   of another strain
     B. Transduction  - genetic transfer among bacteria involving the use of a virus
            as the transmitting agent
          1. plasmids used extensively in DNA research; piece of DNA incorporated into existing
                DNA 

 
 Gene Expression/ Regulation  (text pg 152-157)

        OPERON CONCEPT- (Objective #17)  part of each gene acts as a "switch" to either
               turn "on" or "off" depending on kind of cell or concentrations of gene products
               (proteins)

               *lac operon; pancreas- insulin

            Promoter
            Protein Coding Region
            Termination Sequence
             Regulatory regions


Using DNA to our Advantage  (chapter 11 text pgs 215-236)

     DNA tech.; Gene splicing; Recombinant DNA/ Transgenics
     Human insulin: old ---> pancreas of cattle/sheep; new ---> bacterial
     Gene Therapy
      Forensics


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                                    Use of DNA in Identification
                                    Genetically Modified Organisms
                                    To Know Ourselves
                                    The Gene Hunters
                                    Harvest of Fear


CHROMOSOMES- (Objective #21)  composed of genes; genes- ~450-1000

        nucleotides (DNA)

     gene --> mRNA --> protein --> gene expression
          Specific location on a chromosome referred to as a gene locus

     Chromosome theory- (Objective #22)  since there are more traits than there are
          chromosomes, each chromosome must carry many traits (genes)

          **all cells contain an identical copy of every chromosome (w/exception of egg
                and sperm which have 1/2)

          -however, not all cells have all genes "turned on"

(Objective #23)
     Diploid- chromosomes found in pairs (2N; N= chromosomes)
          Human 2N number is 46
          Dog 2N= 78
     Haploid- single set of chromosomes, characteristic of gametes (eggs and sperm)
          Human haploid (1N) number is 23

          Maternal egg             Paternal sperm           Zygote
               1N          +            1N        ------>                2N
          haploid = 23             haploid = 23                 diploid = 46

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