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    • Female Reproductive System
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    • The Male Reproductive System and Male Contraception
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    • In-Vitro Fertilization
    • Infertility
    • Genetics of Reproduction
    • Prenatal and Maternity Care
    • The Pregnant Body
    • fetal development
    • Development of the Nervous System
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    • Postpartum Issues
    • Twins
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    • pH Lab
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    • Volcano Project
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    • Time Management
  • Environmental Science
    • MIDTERM 2 STUDY GUIDE
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    • ENVS 105 Home Page
      • Midterm 3 Study Guide Population Ecology
      • Ecology II - Communities and Ecosystems
      • Module 1 Assignments
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    • Inrtoduction to ENV SCI
    • Historical Perspective of ​Environmental Science
    • Biomes
    • FOOD CHAIN and FOOD WEB
    • Biogeochemical Recycling
    • Evolution - Our Beginning
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    • Evolution: How Populations Change over Time
    • Symbiosis
    • Population Ecology
    • Competition in Nature
    • Herbivory
    • Niches
    • Fossil Fuels
  • Environmental Biology Laboratory
    • SOILS AND GROUNDWATER
    • Ecological Roles of Living Organisms
      • The Basics
      • Bacteria - Ecological Roles
      • Protists - Ecological Roles
      • Fungus - Ecological Roles
      • Plantae and Animalia - Ecological Roles
    • Virtual FIELD TRIP TO THE RIO HONDO COLLEGE ​WILDLIFE SANCTUARY - Adaptations to Dry Climates
    • Microscopic Plant Adaptations
    • Natural Selection
    • GROWTH CURVES
    • SOILS AND GROUNDWATER
    • LC50 and LD50
    • How to Make a Solar Water Heater
    • WATER QUALITY ANALYSIS
  • General Biology
    • Characteristics of Life
    • Chemistry of Life - Inorganic
    • The Chemistry of Cells - ORGANIC
    • Introduction to The Cell
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    • GENERAL BIO 1110L Labs
      • lab 2 - CELLS - BIO 111L
      • lab 3 - DIFFUSION and OSMOSIS - BIO 111L
      • lab 4 - The Circulatory System - BIO 111L
      • lab 6 - Photosynthesis and Cellular Respiration
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      • DNA, GENES AND GENETIC INHERITANCE
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  • Human Biology
    • A History of Human Biology
    • Levels of Organization
    • The Chemistry of Cells - ORGANIC
    • Cells
    • Cartilage SAC
    • BONES AND SKELETAL TISSUES
  • Human Biology Lab
    • Testing for Sugar, Starch and Proteins
    • Osmosis, Diffusion and Filtration
    • buffers
    • OSMOSIS LAB
    • Anatomical Planes
    • Body Cavities and Membranes
    • Anatomical Positions
    • The Appendicular Skeleton
    • The SKULL
    • the Thoracic Cage
    • the vertebral column
  • Human Sexuality
    • Course Information
    • Course Calendar
    • Lesson 1 - Introduction to Human Sexuality
    • Lesson 2 - Genetic Inheritance of Human Sexuality
    • Lesson 3 - The Male Reproductive Tract
    • Lesson 4 - The Female Reproductive Tract
    • Lesson 5 - The Menstrual Cycle
    • Midterm Exam Study Guide
    • Lesson 6 - Fetal Development and Sexual Differentiation
    • Lesson 7 - Disorders of Sexual Development
    • Lesson 8 - Gender Identity and Sexual Attraction
    • Lesson 9 - Fetishism
    • Lesson 10 - Sexuality Throughout the World
    • ​Lesson 11 - Sexuality Through the Ages
    • Lesson 12 - Sexual Harassment, Coercion and Violence
    • Final Exam Study Guide
  • Microbiology PORTAL
    • Microbiology - CPP
      • ​Intro to Microorganisms
      • Diseases
      • EPIDEMIOLOGY
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      • Bacteria versus Archaea
      • Intro. to Bacteria
      • Viruses and Prions
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      • Microbial Nutrition and Growth
        • Nutritional Categories
        • Microbial Metabolism
        • CONTROL OF BACTERIA GROWTH AND ANTIBIOTICS
      • Eukaryotic Organisms
      • Archaeal Diversity
      • Prokaryotic and Eukaryotic Cells
      • Bacteria vs Archaeal Structures
      • Taxonomic Classifications
      • Archaea, Bacteria and Eukaryotic Cells
      • MIC- CPP Course Calendar
    • Cell Theory
    • Chemistry of Life
      • Chemical Bonds
      • Chemical Reactions
    • Biofilms
    • Definition of Terms
  • Microbiology Laboratory
    • Cell Culture and Inoculations
    • aseptic technique
    • WET MOUNT
    • Streak Plate
    • Mannitol salt agar (MSA) Test
    • Eosin Methylene Blue (EMB)
    • Blood Agar
    • Dilution Series and Calculations
    • Phage Plaque Assay
    • MICROBIOLOGY UNKNOWN LAB
    • Microbiology Lab -study guide exam one
    • Ex 2 - Microorganisms
    • EX 3 - aseptic technique
    • Ex 4 - Smear Prep
    • Ex 5 - Simple Stains
    • Ex 6 - Negative Staining
    • Ex 8 - Gram Stain
    • Ex 9 - Acid-Fast Stain
    • Ex 10 - Endospore Stain
    • Ex 11 - Motility Test
    • ex 12 -​ Pure culture technique
    • ex 13 - UV Radiation
    • Ex 14 - Enumeration of Bacteria : Standard Plate Count
    • ex - 15 Effects of Temperature on Growth
    • ex 16 - Hand-washing
    • ex 17 - pH and microbial growth
    • ex 18 - Evaluation of Antiseptics
    • ex 19 - Antibiotic Sensitivity : Kirby-Bauer Method
  • HISTOTECHNOLOGY
  • The Brain
  • The Brain
  • The Structure of DNA
  • Contact
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    • The Carbon Cycle - ACTIVITY
    • Evolution - ACTIVITY
    • The Cell Game
    • SYMBIOSIS ACTIVITY
    • THE LORAX ACTIVITY
    • Brittney the Kidney
    • From Soup to Poop
    • MITOSIS - THE NURSERY RHYME
    • Verne the Sperm and friends
      • Verne the Sperm pg1
        • Verne the Sperm pg2
        • Verne the Sperm pg3
        • Verne the Sperm pg4
        • Verne the Sperm pg5
  • Lab 6 - The Chemistry of Cells
  • A History of Anatomy
  • List of Pages
    • Microscopes
  • Cell Membranes and Osmosis
  • Chemistry of Life
  • Muscle Movements
  • The Muscles of the Head, Trunk and Shoulders
  • The Muscles of the Limbs
  • Nervous Tissue
  • The Brain - Anat and Physiology
  • Instructions for Taking BIO 3070
  • MTH 121 Algebra A - Course Schedule and Info
  • Laboratory Calendar CMC Spring 2019
  • Genetics Lab
  • Chemistry and Conversions Lab
  • Digestion and Enzymes Lab
  • Endocrine and Homeostasis Lab
  • Muscles and Reflexes Lab
  • Sensory Lab
  • Immunohistochemistry
  • Blood Lab
  • Heart Rate, Blood Pressure, Electrocardiogram Lab
  • Respiratory Lab
  • Lab 11 Renal Lab
  • Blood Typing Game
  • Body Systems Interactive
  • Ch 9 - The Central Nervous System
  • Ch 10 - Sensory Systems
  • Neuron Virtual Laboratory
  • Virtual Eye Lab
  • Virtual pH Lab
  • Chemical Bonds Virtual Lab
  • Beer's Law Virtual Lab
  • Build-an-Atom Virtual Lab
  • Diffusion Virtual Lab
  • Ohm's Law Virtual Lab
  • New Page
  • Ch 8 - Nervous System


 
P-Glo
You should have downloaded the PGlo handout. Work in groups of four.
 
The pGLO plasmid is an engineered plasmid used in biotechnology as a vector for creating genetically modified organisms. The plasmid contains several reporter genes, most notably for the green fluorescent protein (GFP) and the ampicillin resistance gene. GFP was isolated from the jelly fish Aequorea victoria. Because it shares a bidirectional promoter with a gene for metabolizing arabinose, the GFP gene is only expressed in the presence of arabinose, which makes the transgenic organism fluoresce under UV light. GFP can be induced in bacteria containing the pGLO plasmid by growing them on +arabinose plates.
 
pGLO is made up of three genes that are joined together using recombinant DNA technology. They are as follows:
-Bla, which codes for the enzyme beta-lactamase giving the transformed bacteria resistance to the beta-lactam family of antibiotics (such as of the penicillin family)
-araC, a promoter region that regulates the expression of GFP (specifically, the GFP gene will be expressed only in the presence of arabinose)
-GFP, the green fluorescent protein
 
P-Glo is a transformation exercise. We briefly talked about plasmids and transformation in lecture. Some organisms have the ability to take up DNA from their environment. We will use E. coli and force it to take up DNA in the form of a plasmid. In E. coli, there is a promoter region for ampicillin resistance. Another promoter region contains the gene for the sugar, arabinose (inducible operon). E. coli requires arabinose to turn the gene off.
 
We have a plasmid which contains the gene from a jellyfish that allows the organism to glow under florescent light. We are going to introduce the gene into the E. coli, and grow them in plates (LB instead of TSA). Two plates will have E. coli which are ampicillin resistant, two plates will have E. coli which are not ampicillin resistant. One of each of those plates will have E. coli which has had the plasmid inserted, the other will not. One of the other two plates (positive; contains the plasmid) will contain arabinose, the other will not. We will be incubating in the presence of ampicillin, and that will select for those organisms which can take up the plasmid.
We will also be incubating for the presence of the sugar, arabinose, because it is required to turn the gene on. If there is no gene for ampicillin resistance or arabinose, it should not be able to glow.
 
Take one colony and place it in one of each of the pink and blue tubes.
The technician will put plasmid in the pink tube. Put both of the tubes into ice for 10 minutes to force the transformation. Each tube also contains calcium chloride, which pokes holes in the outer membrane, allowing the DNA to gain access to the inside of the cell. Now we have to shock the cell to cause the DNA to be taken up. To do this, put them into the water bath at 57 degrees for 50 seconds, then back on ice for 2 minutes. Then collect your tubes and add 250 µl of LB broth to both tubes and incubate at room temperature for 10 mins. This allows for replication of the cell and incorporation of the plasmid. Then pipette 100 µl from the blue tube to the two negative plates (contain no plasmid; one has arabinose and one does not), and pipette 100 µl from the pink tube to the two positive plates (contains plasmid; one has arabinose and one does not). Next lab, we will expose them to UV light to excite the proteins and see which ones show green fluorescence. The only plate that should glow is the one with the plasmid plus arabinose.
 
Plates
LB/amp positive (has plasmid)
LB/amp/arabinose; positive
LB/amp negative (before transformation, they have no amp resistance. Should not glow)
LB negative (just a media like TSA; will grow, but no glow)
 
The promoter region (can turn off and on; is an operon. It is turned off and on by arabinose).
The non-transformed cells do not have the ampicillin resistance gene.
 
 
​​


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      • Lab Quiz 5
    • Anatomy Basics
      • Intro to Anatomy
      • Medical Terminology
      • A History of Anatomy
      • Levels of Organization
      • Anatomical Positions
      • Anatomical Planes
      • Anatomical Regions
      • Body Cavities and Membranes
    • Cells Portal
      • Anatomy of the Cell SAC
      • Membrane Transport
      • The Cell Cycle
      • REGULATION of The Cell Cycle
      • BLOOD CELLS
      • mitosis
    • Tissues Portal SAC
      • The Integumentary System
      • Epithelial Tissues
      • Connective Tissue
      • Muscle Tissue
      • BONES AND SKELETAL TISSUES
      • Cartilage SAC
    • Organ Systems
    • Portal to the Skeletal system
      • The SKULL ANATOMY
      • the Thoracic Cage
      • the vertebral column
      • The Appendicular Skeleton
      • BONES AND SKELETAL TISSUES
      • joints
    • The Muscular System Portal
      • Muscle Tissue
      • Muscles - Intramuscular Injection Sites - WCU
      • Muscles of the Body - Review
    • The Nervous System
      • Introduction to the Nervous System
      • Nervous Tissue
      • The Brain - Anat
      • The Ear - Sensory Organs
      • The Eye - Sensory Organs
    • THE REPRODUCTIVE SYSTEM
    • The Renal System
    • The Respiratory System
    • THE CIRCULATORY SYSTEM PORTAL
      • Intro to the Circulatory System
      • THE HEART
      • HEART DISSECTION PHOTO GALLERY
      • THE VESSELS OF BLOOD CIRCULATION
    • Digestive System
    • Animal Dissection (Virtual)
    • dissection of the fetal pig
  • Physiology
    • Homeostasis - Physio
    • Chemical Reactions - Physio
    • Chemistry of Life - Inorganic - Physio
    • The Chemistry of Cells - ORGANIC - Physio
    • Chemical Bonds - Physio
    • Metabolism - Physio
    • Portal to the Skeletal system
    • Endocrine and Homeostasis physio
    • Muscle Physiology
    • Blood
    • Cardiovascular System
    • Lymphatic System
    • Respiratory System Physiology
    • Renal System
    • Digestive System
    • Reproductive System
  • CMC Physiology Lab
    • Lab 1 - Surface Area to Volume Ratios
    • Lab 2 - Osmosis
    • Lab 4 - Heart Rate and Barometers
    • Lab 5 - Virtual Neuron Lab
    • Case Study One
  • Anat & Physio
    • The Muscular System Portal
    • The Integumentary System a&p
      • The Epidermis
      • The Dermis
      • The Epidermis rio
      • Connective Tissue
  • Biology of Human Pregnancy
    • Course Calendar - BIO 3070
    • Bio of Pregnancy - SYLLABUS
    • Course Information
    • Evolution of Human Pregnancy
    • History of Human Pregnancy
    • Myths of Pregnancy and Fertility
    • Female Reproductive System
    • The Menstrual Cycle
    • The Male Reproductive System and Male Contraception
    • Fertility and Conception
    • In-Vitro Fertilization
    • Infertility
    • Genetics of Reproduction
    • Prenatal and Maternity Care
    • The Pregnant Body
    • fetal development
    • Development of the Nervous System
    • Stages of Labor
    • Postpartum Issues
    • Twins
  • Chemistry
    • pH Lab
    • The Chemistry of Cells - ORGANIC
      • VOLCANO LAB
    • Volcano Project
  • College/Life Skills
    • Online Professionalism
    • Advising Resources
    • INTERVIEW SKILLS AND RESUME WRITING
    • DIVERSITY
    • CAMPUS EVENTS
      • Predation
    • Time Management
  • Environmental Science
    • MIDTERM 2 STUDY GUIDE
    • Exam 2 Study Guide
    • ENVS 105 Home Page
      • Midterm 3 Study Guide Population Ecology
      • Ecology II - Communities and Ecosystems
      • Module 1 Assignments
      • Module 2 Assignments
    • Inrtoduction to ENV SCI
    • Historical Perspective of ​Environmental Science
    • Biomes
    • FOOD CHAIN and FOOD WEB
    • Biogeochemical Recycling
    • Evolution - Our Beginning
    • Genetic Inheritance
    • Evolution: How Populations Change over Time
    • Symbiosis
    • Population Ecology
    • Competition in Nature
    • Herbivory
    • Niches
    • Fossil Fuels
  • Environmental Biology Laboratory
    • SOILS AND GROUNDWATER
    • Ecological Roles of Living Organisms
      • The Basics
      • Bacteria - Ecological Roles
      • Protists - Ecological Roles
      • Fungus - Ecological Roles
      • Plantae and Animalia - Ecological Roles
    • Virtual FIELD TRIP TO THE RIO HONDO COLLEGE ​WILDLIFE SANCTUARY - Adaptations to Dry Climates
    • Microscopic Plant Adaptations
    • Natural Selection
    • GROWTH CURVES
    • SOILS AND GROUNDWATER
    • LC50 and LD50
    • How to Make a Solar Water Heater
    • WATER QUALITY ANALYSIS
  • General Biology
    • Characteristics of Life
    • Chemistry of Life - Inorganic
    • The Chemistry of Cells - ORGANIC
    • Introduction to The Cell
    • Photosynthesis and cellular Respiration
    • Cell Membranes and Osmosis
    • The Cell Cycle
    • REGULATION of The Cell Cycle
    • Mitosis
    • Meiosis
    • The Structure of DNA
    • Evolution
  • General Biology Laboratory
    • GENERAL BIOLOGY 101 LABORATORY HOME PAGE
      • Enzymes
      • OSMOSIS LAB
      • Lab 1 - Bacteria, Protista and Fungi
      • Lab 2 - Plantae and Animalia
      • Photosynthesis
      • Lab 5 - Introduction to Cells
      • Lab 6 - The Chemistry of Cells
      • Lab 7 - Membrane Transport
      • Lab 8 - Enzymes
      • Lab 9 - Photosynthesis
      • Lab 10 Fermentation, Aerobic Cellular Respiration and Associated Major Organ Systems
    • GENERAL BIO 1110L Labs
      • lab 2 - CELLS - BIO 111L
      • lab 3 - DIFFUSION and OSMOSIS - BIO 111L
      • lab 4 - The Circulatory System - BIO 111L
      • lab 6 - Photosynthesis and Cellular Respiration
      • lab 7 - Reproduction - BIO 111L
      • DNA, GENES AND GENETIC INHERITANCE
      • lab 9 - GENE EXPRESSION AND PROTEIN SYNTHESIS
      • lab 10 - ADAPTATIONS - BIO 111L
      • lab 11 - ECOSYSTEMS AND BIODIVERSITY
  • Human Biology
    • A History of Human Biology
    • Levels of Organization
    • The Chemistry of Cells - ORGANIC
    • Cells
    • Cartilage SAC
    • BONES AND SKELETAL TISSUES
  • Human Biology Lab
    • Testing for Sugar, Starch and Proteins
    • Osmosis, Diffusion and Filtration
    • buffers
    • OSMOSIS LAB
    • Anatomical Planes
    • Body Cavities and Membranes
    • Anatomical Positions
    • The Appendicular Skeleton
    • The SKULL
    • the Thoracic Cage
    • the vertebral column
  • Human Sexuality
    • Course Information
    • Course Calendar
    • Lesson 1 - Introduction to Human Sexuality
    • Lesson 2 - Genetic Inheritance of Human Sexuality
    • Lesson 3 - The Male Reproductive Tract
    • Lesson 4 - The Female Reproductive Tract
    • Lesson 5 - The Menstrual Cycle
    • Midterm Exam Study Guide
    • Lesson 6 - Fetal Development and Sexual Differentiation
    • Lesson 7 - Disorders of Sexual Development
    • Lesson 8 - Gender Identity and Sexual Attraction
    • Lesson 9 - Fetishism
    • Lesson 10 - Sexuality Throughout the World
    • ​Lesson 11 - Sexuality Through the Ages
    • Lesson 12 - Sexual Harassment, Coercion and Violence
    • Final Exam Study Guide
  • Microbiology PORTAL
    • Microbiology - CPP
      • ​Intro to Microorganisms
      • Diseases
      • EPIDEMIOLOGY
      • HOST DEFENSES
      • PATHOGENICITY
      • History of Microbiology
      • Levels of Organization cpp
      • Bacteria versus Archaea
      • Intro. to Bacteria
      • Viruses and Prions
      • Microbial Genetics
      • Microbial Nutrition and Growth
        • Nutritional Categories
        • Microbial Metabolism
        • CONTROL OF BACTERIA GROWTH AND ANTIBIOTICS
      • Eukaryotic Organisms
      • Archaeal Diversity
      • Prokaryotic and Eukaryotic Cells
      • Bacteria vs Archaeal Structures
      • Taxonomic Classifications
      • Archaea, Bacteria and Eukaryotic Cells
      • MIC- CPP Course Calendar
    • Cell Theory
    • Chemistry of Life
      • Chemical Bonds
      • Chemical Reactions
    • Biofilms
    • Definition of Terms
  • Microbiology Laboratory
    • Cell Culture and Inoculations
    • aseptic technique
    • WET MOUNT
    • Streak Plate
    • Mannitol salt agar (MSA) Test
    • Eosin Methylene Blue (EMB)
    • Blood Agar
    • Dilution Series and Calculations
    • Phage Plaque Assay
    • MICROBIOLOGY UNKNOWN LAB
    • Microbiology Lab -study guide exam one
    • Ex 2 - Microorganisms
    • EX 3 - aseptic technique
    • Ex 4 - Smear Prep
    • Ex 5 - Simple Stains
    • Ex 6 - Negative Staining
    • Ex 8 - Gram Stain
    • Ex 9 - Acid-Fast Stain
    • Ex 10 - Endospore Stain
    • Ex 11 - Motility Test
    • ex 12 -​ Pure culture technique
    • ex 13 - UV Radiation
    • Ex 14 - Enumeration of Bacteria : Standard Plate Count
    • ex - 15 Effects of Temperature on Growth
    • ex 16 - Hand-washing
    • ex 17 - pH and microbial growth
    • ex 18 - Evaluation of Antiseptics
    • ex 19 - Antibiotic Sensitivity : Kirby-Bauer Method
  • HISTOTECHNOLOGY
  • The Brain
  • The Brain
  • The Structure of DNA
  • Contact
  • FUN ZONE
    • GAMES
    • Video Vault
    • Population Ecology - ACTIVITY
    • The Carbon Cycle - ACTIVITY
    • Evolution - ACTIVITY
    • The Cell Game
    • SYMBIOSIS ACTIVITY
    • THE LORAX ACTIVITY
    • Brittney the Kidney
    • From Soup to Poop
    • MITOSIS - THE NURSERY RHYME
    • Verne the Sperm and friends
      • Verne the Sperm pg1
        • Verne the Sperm pg2
        • Verne the Sperm pg3
        • Verne the Sperm pg4
        • Verne the Sperm pg5
  • Lab 6 - The Chemistry of Cells
  • A History of Anatomy
  • List of Pages
    • Microscopes
  • Cell Membranes and Osmosis
  • Chemistry of Life
  • Muscle Movements
  • The Muscles of the Head, Trunk and Shoulders
  • The Muscles of the Limbs
  • Nervous Tissue
  • The Brain - Anat and Physiology
  • Instructions for Taking BIO 3070
  • MTH 121 Algebra A - Course Schedule and Info
  • Laboratory Calendar CMC Spring 2019
  • Genetics Lab
  • Chemistry and Conversions Lab
  • Digestion and Enzymes Lab
  • Endocrine and Homeostasis Lab
  • Muscles and Reflexes Lab
  • Sensory Lab
  • Immunohistochemistry
  • Blood Lab
  • Heart Rate, Blood Pressure, Electrocardiogram Lab
  • Respiratory Lab
  • Lab 11 Renal Lab
  • Blood Typing Game
  • Body Systems Interactive
  • Ch 9 - The Central Nervous System
  • Ch 10 - Sensory Systems
  • Neuron Virtual Laboratory
  • Virtual Eye Lab
  • Virtual pH Lab
  • Chemical Bonds Virtual Lab
  • Beer's Law Virtual Lab
  • Build-an-Atom Virtual Lab
  • Diffusion Virtual Lab
  • Ohm's Law Virtual Lab
  • New Page
  • Ch 8 - Nervous System