Lecture PowerPoint to accompany Inquiry into Life Twelfth Edition Sylvia S. Mader Chapter 24 Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display. 24.1 DNA Structure and Replication • Hershey-Chase Experiments (1952) – Demonstrated that DNA is the genetic material – DNA stands for Deoxyribonucleic Acid Hershey-Chase Experiments 24.1 DNA Structure and Replication • Structure of DNA – James Watson and Francis Crick determined the structure of DNA in 1953 – DNA is a chain of nucleotides – Each nucleotide is a complex of three subunits • Phosphoric acid (phosphate) • A pentose sugar (deoxyribose) • A nitrogen-containing base 24.1 DNA Structure and Replication • Structure of DNA – Four Possible Bases • Adenine (A) - a purine • Guanine (G) - a purine • Thymine (T) - a pyrimidine • Cytosine (C) - a pyrimidine – Complimentary Base Pairing • Adenine (A) always pairs with Thymine (T) • Guanine (G) always pairs with Cytosine (C) Overview of DNA Structure 24.1 DNA Structure and Replication • Replication of DNA – Semi-conservative replication • Each daughter DNA molecule consists of one new chain of nucleotides and one from the parent DNA molecule – The two daughter DNA molecules will be identical to the parent molecule 24.1 DNA Structure and Replication • Replication of DNA – Before replication begins, the two strands of the parent molecule are hydrogen-bonded together – Enzyme DNA helicase unwinds and “unzips” the doublestranded DNA – New complementary DNA nucleotides fit into place along divided strands by complementary base pairing. These are positioned and joined by DNA polymerase – DNA ligase repairs any breaks in the sugar-phosphate backbone – The Two double helix molecules identical to each other and to the original DNA molecule Overview of DNA Replication Ladder Configuration and DNA Replication 24.2 Gene Expression • Gene: A segment of DNA that specifies the amino acid sequence of a polypeptide • DNA does not directly control protein synthesis, instead its information is transcribed into RNA 24.2 Gene Expression • RNA (ribonucleic acid) 24.2 Gene Expression • Three Classes of RNA – Messenger RNA (mRNA) • Takes a message from DNA to the ribosomes – Ribosomal RNA (rRNA) • Makes up ribosomes (along with proteins) – Transfer RNA (tRNA) • Transfers amino acids to ribosomes 24.2 Gene Expression • Gene Expression Requires Two Steps: – Transcription – Translation 24.2 Gene Expression • Transcription – During transcription, a segment of the DNA serves as a template for the production of an RNA molecule – Messenger RNA (mRNA) • RNA polymerase binds to a promoter • DNA helix is opened so complementary base pairing can occur • RNA polymerase joins new RNA nucleotides in a sequence complementary to that on the DNA 24.2 Gene Expression • Transcription – Processing of mRNA • Primary mRNA becomes mature mRNA • Contains bases complementary to both intron and exon segments of DNA – Introns are intragene segments – Exons are the portion of a gene that is expressed • Intron sequences are removed, and a poly-A tail is added – Ribozyme splices exon segments together Transcription of DNA to form mRNA mRNA Processing 24.2 Gene Expression • Translation – The Genetic Code • Triplet code- each 3-nucleotide unit of a mRNA molecule is called a codon • There are 64 different mRNA codons – 61 code for particular amino acids » Redundant code-some amino acids have numerous code words » Provides some protection against mutations – 3 are stop codons signal polypeptide termination Messenger RNA Codons 24.2 Gene Expression • Transfer RNA – tRNA transports amino acids to the ribosomes – Single stranded nucleic acid that correlates a specific nucleotide sequence with a specific amino acid – Amino acid binds to one end, the opposite end has an anticodon – the order of mRNA codons determines the order in which tRNA brings in amino acids Transfer RNA: Amino Acid Carrier 24.2 Gene Expression • Ribosome and Ribosomal RNA – Ribosome has a binding site for mRNA and for 2 tRNAs – Facilitate complementary base pairing – Ribosome moves along mRNA and new tRNAs come in and line up in order – This brings amino acids in line in a specific order to form a polypeptide – Several ribosomes may move along the same mRNA • Multiple copies of a polypeptide may be made • The entire complex is called a polyribosome Polyribosome Structure and Function Overview of Gene Expression 24.2 Gene Expression • Translation Requires Three Steps – Initiation (requires energy) – Elongation (requires energy) – Termination Initiation Elongation Termination Summary of Gene Expression 24.2 Gene Expression • Genes and Gene Mutations – A gene mutation is a change in the sequence of bases within a gene. – Gene mutations can lead to malfunctioning proteins in cells. 24.2 Gene Expression • Genes and Gene Mutations – Causes of Mutations • Errors in replication – Rare – DNA polymerase “proofreads” new strands and errors are cleaved out • Mutagens – Environmental influences – Radiation, UV light, chemicals – Rate is low because DNA repair enzymes monitor and repair DNA • Transposons – “jumping genes” – Can move to new locations and disrupt sequences Transposon 24.2 Gene Expression • Types of Mutations – Frameshift Mutations • One or more nucleotides are inserted or deleted • Results in a polypeptide that codes for the wrong sequence of amino acids – Point Mutations • The substitution of one nucleotide for another – Silent mutations – Nonsense mutations – Missense mutations 24.3 DNA Technology • The Cloning of a Gene – Cloning: Production of many identical copies of an organism through some asexual means. – Gene Cloning: The production of many identical copies of a single gene – Two Ways to Clone a Gene: – Recombinant DNA – Polymerase Chain Reaction Cloning of a Human Gene / Recombinant DNA 24.3 DNA Technology • Using Recombinant DNA Technology – Restriction enzymes breaks open a plasmid vector at specific sequence of bases “sticky ends” – Foreign DNA that is to be inserted is also cleaved with same restriction enzyme so ends match – Foreign DNA is inserted into plasmid DNA and “sticky ends” pair up – DNA ligase seals them together Restriction Enzymes and Stick Ends 24.3 DNA Technology • Polymerase Chain Reaction – Amplifies a targeted DNA sequence – Requires DNA polymerase, a set of primers, and a supply of nucleotides • Primers are single stranded DNA sequences that start replication process – Amount of DNA doubles with each replication cycle – Process is now automated Polymerase Chain Reaction (PCR) (insert figure 24.18) 24.3 DNA Technology • DNA Fingerprinting – Permits identification of individuals and their relatives – Based on differences between sequences in nucleotides between individuals – Detection of the number of repeating segments (called repeats) are present at specific locations in DNA • Different numbers in different people • PCR amplifies only particular portions of the DNA • Procedure is performed at several locations to identify repeats DNA Fingerprints 24.3 DNA Technology • Biotechnology – Biotechnology uses natural biological systems to create a product or to achieve a goal desired by humans. – Transgenic organisms have a foreign gene inserted into their DNA 24.3 DNA Technology • Transgenic Bacteria – Medical Uses: Production of Insulin, Human Growth Hormone, Tissue Plasminogen Activator, Hepatitis B Vaccine – Agricultural Uses: Bacteria that protects plants from freezing, bacteria that protect plant roots from insects – Environmental: Bacteria that degrade oil (clean up after oil spills), bacteria that remove sulfur from coal 24.3 DNA Technology • Transgenic Plants – Plants have been engineered to secrete a toxin that kills insects – Plants have been engineered to be resistant to herbicides 24.3 DNA Technology • Transgenic Animals – Fish, cows, pigs, rabbits and sheep have been engineered to produce human growth hormone in order to increase size of the animals Transgenic Animals