BIOL 2051 Lecture Notes - Lecture 6: Citrobacter Freundii, Staphylococcus Epidermidis, Lactococcus Lactis

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29 Nov 2016

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There will be 3 parts to the final exam, all done on the last day of lab. Section a you perform an experiment and interpret the results. With knowledge of experiments 21-37, you will be able to perform the experiment and interpret the results. Section b questions about a set of materials (tubes, plates, etc) on the bench top in front of you. Section c short answer/open response questions: general information: Materials to study: background material and procedures in lab manual. Phage titration problem: there will be at least one phage titration problem on the midterm. Organisms you need to know: know what is unique about each & test(s) to distinguish them. 34 capable of causing severe illness, especially in patients who are hospitalized. Produces and enterotoxin which is responsible for sever gastroenteritis or staphylococcal food poisoning. Produces a toxin that causes toxic shock syndrome.

Related Questions

Biology lab homework

Dragon Genes

http://www.concord.org/stem-resources/modern-genetics

BLY 121L - Take HomeAssignment

Due: Start of class time âAt thestart of class next week

Answers MUST beâ¦

â¢ typed (no smaller than 11-pointfont)

â¢ complete sentences (i.e., grammaticallycorrect â AND spell check!)

â¢ THIS IS NOT AGROUP EFFORT â NO POINTS WILL BE AWARDED, IF YOUR PAPER ISSIMILAR IN CONTENT AND WORDING TO THAT OF YOURPEERâS.

Complete ALL4 modules:

1. Intro& Rules

2. Meiosis

3. Pedigree andGenes

4. The PlatesProblem

Complete and answer all of the questions fromeach module.

Introduction

Breed dragons to yield several offspring anddevelop a chart for all of the phenotypic characteristics youobserve.

â¢ Note the genotype and dominant/recessivecondition for each trait.

â¢ When you reach this section of theexercise, be sure to change the genotype for all traits to completethe chart.

Whatare the traits exhibited by yourdragons?

**YOUMUST TURN IN YOURCOMPLETED CHART AND IT MUST BE LEGIBLE!

Rules

Dominance and Recessive Traits

What isthe chromosome number(ânâ) for thedragons?

Which trait(s)exhibit incompletedominance?

What does it mean fora trait to be incompletely dominant?

Noallelic combination is described as being heterozygousrecessive. Why do you thinkthis is thecase?

Some Traits are Sex-linked

Which dragontrait(s)is/are sex-linked?

How dosex-linked traits differ from autosomal traits?

Color and Lethal Combinations

Describehow the inheritance ofcolor is different from that of horns and/orwings.

On whichchromosome is the lethal combination exhibited? What is the lethalcombination?

Meiosis

What is the phenotypeof your new dragon?

Whichtraits areinherited from the mother dragon? AND Which traits are inherited from the father dragon?

Wereany traits expressed by the offspring that differed from either or both of the parents? If so, whatwere they?

How does meiosisallow for variation inoffspring?

How doesfertilization allow for variation in offspring?

Pedigree and Genes

Now that you know how to breed dragons, let'stry some selective breeding in the pedigree view. Use the cross tool tobreed a new generation of dragons; breed two of thedragons from this new (F1) generation. Repeat thisat least8 times.

What do younotice about the dragons as you continue to breed more generations?

Use the chromosome tool to closelyexaminethe dragonsâ genes. If you had trouble answering the sex-linkedquestion above, now you can observe and compare the Xand Y chromosomesâ¦

Can you find any genesthat are different among the members of your F8generation?

How many generationsdo you have to breed until all the dragons are exactly thesame?

You arecreating an inbred strain of dragons in the final generation(s)of this process. Write a notedescribing your strain, and explain what is âspecialâ aboutall individuals in an inbred strain of organisms.

The Plates Problem

Try breedingsome offspring to see resultsof these crosses. Does thepattern of inheritance ofplates tell you anything useful?

Based on yourbreeding experiments, where might you expectto find the gene forplates (i.e., On whichchromosome might the gene be located?) Why do you think this iscorrect?

Dragons thatgrow plates usually do so quite late in their life, so dragons thathave a new baby don't know right away whether or not it will growplates. Which trait(s)exhibited by a baby might helpa mother dragon determine whether her baby will growup to have plates?

DragonGenes
Genotypes/Phenotypes

Trait	Phenotype	Genotype
Horns	Horns
	No Horns
Wings	Wings
	No Wings
Legs	4 Legs
	2 Legs
	No Legs
Tails	Fancy Tail
	Plain Tail
Fire-breathing	Fire-breathing Male
	Non-fire-breathing Male
	Fire-breathing Female
	Non-fire-breathingFemale
Color	Green Male
	Yellow Male
	Red Female
	Blue Female
	Brown Female
	Purple Female

olivedonkey40

HELP ASAP;

Please help me understand my lab results, so be detailed to clearly understand . INclude answers to these questions what role does ampicilin (amp) and arabinose (ara) play in the experiment. Because GFP was isolated from jelly fisht, what modification to the GFP DNA had to be made to result in expression in E. coli?

BACKGROUND INFORMATION

Transformation of GFP and the pGLO Plasmid

GFP is a commonly used reporter protein in research labs, as the fluorescence creates a marker protein that can be used in many types of cell biology and biochemical studies. In basic research, GFP is often fused to a specific target protein of interest, creating a chimeric reporter protein. GFP has been used as a reporter protein to study blood vessel and tumor progression in mice, brain activity in mice, and malaria eradication in mosquitoes for instance (see website mentioned in notes).

In the first week you will use a procedure to genetically transform bacteria with the gene that codes for Green Fluorescent Protein. Following the transformation procedure, the bacteria express their newly acquired jellyfish gene and produce the fluorescent protein, which causes them to glow a brilliant green color under ultraviolet light. During the second week you will purify the protein away from all of the other proteins produced by the bacteria using chromatography, and during the third week you will evaluate the success of the purification procedure and estimate the size (molecular weight) of GFP by using sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE).

Fundamental to the process of genetic transformation of bacteria are plasmids. In addition to one large chromosome, bacteria naturally contain one or more small circular pieces of DNA called plasmids. Plasmid DNA usually contains genes for one or more traits that may be beneficial to bacterial survival. In nature, bacteria can transfer plasmids back and forth allowing them to share these beneficial genes. This natural mechanism allows bacteria to adapt to new environments. The occurrence of bacterial resistance to antibiotics is due in large part to the transmission of plasmids which contain antibiotic resistance genes.

The pGLO plasmid (Fig. 5, not a naturally occurring plasmid) contains the gene for GFP, and also contains the gene for the enzyme Î²-lactamase (bla), which provides resistance to the antibiotic ampicillin. The Î²-lactamase protein is produced and secreted by bacteria that contain the plasmid. Î²-Lactamase inactivates the ampicillin present in the LB nutrient agar, allowing bacterial growth. Only transformed bacteria that contain the plasmid and express Î²-lactamase can survive on plates that contain ampicillin. Only a very small percentage of the cells take up the plasmid DNA and are transformed. Untransformed cells cannot grow on the ampicillin selection plates. pGLO also incorporates a special gene regulation system, which can be used to control expression of the fluorescent protein in transformed cells. The gene for GFP can be switched on in transformed cells by adding the sugar arabinose to the cellsâ nutrient medium. Selection for cells that have been transformed with pGLO DNA is accomplished by growth on antibiotic plates. Transformed cells will appear white (wild-type phenotype) on plates not containing arabinose, and fluorescent green when arabinose is included in the nutrient agar medium.

for the first week section of lab

To each of two microfuge tubes, add 250ul of transformation buffer. Keep these tubes on ice.

Using a sterile loop, pick one colony of bacteria from the LB plate.

Transfer this colony to one of the microfuge tubes. Gently vortex the tube until the colony is dispersed. Repeat for the second tube.

Add 0.08ug of DNA to one of these tubes (+DNA tube).

Incubate both tubes on ice for 10 minutes.

Incubate both tubes at 42Â°C for 50 seconds.

Incubate both tubes on ice for 2 minutes.

Add 250ul of LB broth to each tube and incubate at room temperature for 10 minutes.

Flick each tube gently. Add 100ul of this transformation mix to the appropriate plates.

Results:

+DNA: LB/amp--> 1 colony, Glow: uncertain

+DNA: LB/amp/ara--> 9 colonies, Glow: yes

-DNA: LB/amp--> 7 colonies, GLow: no

-DNA: LB--> many colonies, Glow: no

HELP ASAP; Please help me understand my lab results, so be detailed to clearly understand

Then , Please include answers to the question below to include in your response

1. what role does ampicilin (amp) and arabinose (ara) play in the experiment.

2) Because GFP was isolated from jelly fisht, what modification to the GFP DNA had to be made to result in expression in E. coli?

BACKGROUND INFORMATION

Transformation of GFP and the pGLO Plasmid GFP is a commonly used reporter protein in research labs, as the fluorescence creates a marker protein that can be used in many types of cell biology and biochemical studies. In basic research, GFP is often fused to a specific target protein of interest, creating a chimeric reporter protein. GFP has been used as a reporter protein to study blood vessel and tumor progression in mice, brain activity in mice, and malaria eradication in mosquitoes for instance (see website mentioned in notes). In the first week you will use a procedure to genetically transform bacteria with the gene that codes for Green Fluorescent Protein. Following the transformation procedure, the bacteria express their newly acquired jellyfish gene and produce the fluorescent protein, which causes them to glow a brilliant green color under ultraviolet light. During the second week you will purify the protein away from all of the other proteins produced by the bacteria using chromatography, and during the third week you will evaluate the success of the purification procedure and estimate the size (molecular weight) of GFP by using sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). Fundamental to the process of genetic transformation of bacteria are plasmids. In addition to one large chromosome, bacteria naturally contain one or more small circular pieces of DNA called plasmids. Plasmid DNA usually contains genes for one or more traits that may be beneficial to bacterial survival. In nature, bacteria can transfer plasmids back and forth allowing them to share these beneficial genes. This natural mechanism allows bacteria to adapt to new environments. The occurrence of bacterial resistance to antibiotics is due in large part to the transmission of plasmids which contain antibiotic resistance genes. The pGLO plasmid contains the gene for GFP, and also contains the gene for the enzyme Î²-lactamase (bla), which provides resistance to the antibiotic ampicillin. The Î²-lactamase protein is produced and secreted by bacteria that contain the plasmid. Î²-Lactamase inactivates the ampicillin present in the LB nutrient agar, allowing bacterial growth. Only transformed bacteria that contain the plasmid and express Î²-lactamase can survive on plates that contain ampicillin. Only a very small percentage of the cells take up the plasmid DNA and are transformed. Untransformed cells cannot grow on the ampicillin selection plates. pGLO also incorporates a special gene regulation system, which can be used to control expression of the fluorescent protein in transformed cells. The gene for GFP can be switched on in transformed cells by adding the sugar arabinose to the cellsâ nutrient medium. Selection for cells that have been transformed with pGLO DNA is accomplished by growth on antibiotic plates. Transformed cells will appear white (wild-type phenotype) on plates not containing arabinose, and fluorescent green when arabinose is included in the nutrient agar medium.

The transformation procedure involves three main steps. These steps are intended to introduce the plasmid DNA into the E. coli cells and provide an environment for the cells to express their newly acquired genes. 1) Cells are first treated with a solution of CaCl2, which is thought to neutralize the repulsive negative charges of the phosphate backbone of DNA and the phospholipids of the cell membrane, allowing the DNA to adhere to the cells. This is referred to making the cells âcompetentâ (i.e. the cells are capable, or competent, to take-in exogenous DNA). 2) Cells are then subjected to a heat shock, which is thought to increase the permeability of the cell membrane, allowing the cells to take-in the plasmid DNA. 3) Cells are allowed to grow in a ârecoveryâ phase in the absence of ampicillin. During this time the cells begin to produce the Î²-lactamase protein, which will allow them to survive when they are subsequently placed on agar plates containing ampicillin.

For the first week section of lab

To eac

h of two microfuge tubes, add 250ul of transformation buffer. Keep these tubes on ice.

Using a sterile loop, pick one colony of bacteria from the LB plate. Transfer this colony to one of the microfuge tubes.

Gently vortex the tube until the colony is dispersed. Repeat for the second tube. Add 0.08ug of DNA to one of these tubes (+DNA tube).

Incubate both tubes on ice for 10 minutes. Incubate both tubes at 42Â°C for 50 seconds.

Incubate both tubes on ice for 2 minutes. Add 250ul of LB broth to each tube and incubate at room temperature for 10 minutes. Flick each tube gently. Add 100ul of t

his transformation mix to the appropriate plates.

Results For plates:

+DNA: Has the E. Coli pGlo plasmid and - DNA plates do no have EColi pGLo plasmid .

Amp means ampiclin and Ara means arabinose on agar plates

+ DNA Plate LB/amp/ara--> white 9 colonies, Glow: yes

+DNA LB/amp--> 7 colonies, GLow: no

--DNA LB/AMP- 1 colony so no colony growth and no glovw

-DNA: LB--> many colonies, Glow: no

Please explain significance of result or thand the reason for the results in scientific terms based on the background information provided and your own knowledge to be detailed. Would these results be expected as wll.

BIOL 2051 Lecture Notes - Lecture 6: Citrobacter Freundii, Staphylococcus Epidermidis, Lactococcus Lactis

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