MICB 201 Chapter Notes - Chapter 13: Autoclave, Thermophile, Catalase

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copperswan658

15 Dec 2017

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Taking organisms out of their environment will lead to death. Not lowering to the point where water freezes (there is some liquid) Very hard to study (because we have to keep water liquid and not freeze) Have to add solvents to water to keep water from freezing. There is very little molecular motion going on in the gel state. The bilayer is thinner in the fluid state. When the temperature is high enough, you can lose the molecular structure. Melting: gel state heat fluid state (not really from solid to liquid) Gelling: fluid state gelling gel state. The maximum temperature is affected by the dissolving of the membrane and protein denaturation. Permeases bind to nutrients in the environment and transport them into the cytoplasm of the organism. They have to undergo conformational changes in order to change it. If the membrane in the cell is rigid, the conformation is hindered and not able to transport these proteins through the membrane.

Related Questions

I need help on this microbiology question:

Contrary to our favorite science fiction books and movies, the chances of us finding another planet with a civilization comparable to, or even more advanced than our own, seems highly unlikely. But, through your work as a microbiologist at NASA, you have been able to help change our views as to what forms of life we should be looking for on planets in our solar system and around neighboring stars. Instead of radio signals or errant transmissions, we have begun looking for planets that have environments similar to those capable of sustaining life as we know it here on Earth. To determine what we could find out there, you have begun studying different unique and atypical environments found here on Earth. Although the harsh conditions of hot springs or undersea sulfur vents seem incompatible with most forms of life, organisms known collectively as "extremophiles" have still managed to survive and adapt to these extreme conditions. You believe extremophiles may help you gain insight into the potential metabolic requirements, processes, and challenges of extraterrestrial life. Thus far in your search, you have identified some prokaryotic cells that have evolved to survive in the extreme conditions of sulfur vents on the sea floor. These vents are found at great depths, where the crushing pressure and lack of light and oxygen are prohibitive to many forms of life. Bacteria that are not considered extremophiles reside in most ecological niches on this planet. There are some common criteria for the environment that would sustain these bacteria. Availability of water, pH that lies between 5 and 9, temperatures that will remain steady and below boiling; these are some of the characteristics that bacteria will require in their environment if they are to succeed. But in the environment of the ocean floor vents, not only is the pH of the water very low, but also the water temperature is unusually high due to the geologic activity that fuels the vents. Organisms that are adapted to such harsh conditions have necessarily evolved enzymes able to function beyond average optima, as well as a chemosynthetic metabolism capable of using vent organic and inorganic compounds as energy sources.

Part A - Comparable Earth Environments to Be Found on Another Planet

You identify the prokaryotic cells you've found as a species of bacteria. These bacteria appear to be living off of nutrients and sulfur compounds released from the vents on the sea floor. Over time, the bacteria must have evolved extreme metabolic and enzymatic capabilities to allow them to tolerate and even benefit from the extreme depths, the lack of oxygen, the low pH of the surrounding waters, high temperatures due to volcanic releases, and the unique nutrient source of the vents.

Choose the characteristics and their impact on enzyme activity that are not correct from the following statements. (more than one may be chosen)

Archaea are able to tolerate highly acidic pH environments and have proteins that will not denature in the low-pH environments that the bacteria are found in.

Archaea are able to colonize water that is at or above the boiling temperature due to high levels of disulfide bridges that maintain the three-dimensional structure of the enzyme.

Archaea are only able to survive exceedingly well at high temperatures due to the amount of activation energy required by their enzymes.

Archaea are able to grow in extremely shady or low-light conditions that allow for optimal enzyme activity.

champagneworm170

Question 55.55. The fact that marsupials are not as diverse inSouth America as they are in Australia would be classified as whattype of evidence for evolution? (Points : 1.25)

biogeographical

anatomical

biochemical

fossil record

molecular evidence

Question 56.56. What is the name of the hypothesis that statesthat the very first living thing came from nonliving chemicals?(Points : 1.25)

cell theory

chemical evolution

Darwinian evolution

speciation

organic chemistry

Question 57.57. What structures are necessary for thereplication process to occur in a cell? (Points : 1.25)

enzymatic proteins

carbohydrates

lipids

starches

viruses

Question 58.58. The flipper of a whale and the wing of a birdare homologous structures. (Points : 1.25)

TRUE

FALSE

Question 59.59. Both the RNA-first and the protein-firsthypotheses must propose a manner for DNA to become the geneticmaterial as it is today. (Points : 1.25)

TRUE

FALSE

Question 60.60. The early atmosphere of Earth consisted mainlyof (Points : 1.25)

hydrogen, oxygen, and carbon dioxide.

oxygen, water vapor, and carbon monoxide.

water vapor, nitrogen, and carbon dioxide.

hydrogen, carbon monoxide, and nitrogen.

water vapor, hydrogen, and carbon dioxide.

Question 61.61. Which of the following biogeochemical cycles isa sedimentary cycle? (Points : 1.25)

carbon

nitrogen

phosphorus

water

ice

Question 62.62. What two characteristics define a biome? (Points: 1.25)

temperature and the types of plants found there

amount of precipitation and the types of plantsfound there

temperature and rainfall

latitude and rainfall

altitude and temperature

Question 63.63. The atmosphere would be considered whatcomponent of a chemical cycle? (Points : 1.25)

reservoir

exchange pool

biotic community

source

none of these

Question 64.64. Are humans capable of upsetting globalbiogeochemical cycles with their activities? (Points : 1.25)

YES

Question 65.65. The richest marine ecosystems are those in theopen water where large fish abound. (Points : 1.25)

TRUE

FALSE

Question 66.66. What is involved in the process ofdenitrification? (Points : 1.25)

conversion of nitrogen gas to ammonium

conversion of nitrogen gas to nitrate

conversion of ammonium to nitrate

conversion of nitrite to nitrate

conversion of nitrate to nitrogen gas

Question 67.67. What is the loss of water from plants called?(Points : 1.25)

runoff

precipitation

evaporation

transpiration

transference

Question 68.68. Decomposers perform a valuable service becausethey recycle nutrients. (Points : 1.25)

TRUE

FALSE

Question 69.69. There is more nitrogen gas in the atmospherethan there is oxygen. (Points : 1.25)

TRUE

FALSE

Question 70.70. Which of the following defines an ecosystems?(Points : 1.25)

a group of organisms of the same speciesinteracting together

all of the living organisms in an area interactingtogether

all living organisms and the nonliving environmentinteracting together

aspects of the nonliving environment interactingtogether

all the places on Earth where living organisms canbe found

Question 71.71. In MDCs, many people do not eat enough protein.(Points : 1.25)

TRUE

FALSE

Question 72.72. The age structure diagram of MDCs is shaped like(Points : 1.25)

a pyramid.

an inverted pyramid.

a rectangle.

an oval.

a cube.

Question 73.73. The biggest problem in moving toward asustainable society with the LDCs is overconsumption. (Points :1.25)

TRUE

FALSE

Question 74.74. A sustainable society is one that provides allof the goods and services desired by humans. (Points : 1.25)

TRUE

FALSE

Question 75.75. In order to have a negative population growthrate, (Points : 1.25)

the birthrate must exceed the death rate.

the biotic potential of the population mustincrease.

the death rate must exceed the birth rate.

the carrying capacity for the population mustincrease.

only replacement reproduction can occur.

Question 76.76. Why should pregnant women and small children noteat certain fish? (Points : 1.25)

herbicide contamination

mercury contamination

pesticide contamination

lead contamination

PCB contamination

Question 77.77. What do hydropower and wind power have incommon? (Points : 1.25)

Both require the building of dams.

Both lead to an increase in greenhouse gasemissions.

Both are renewable energy sources.

Both result in the depletion of fossil fuels.

Both require the use of photovoltaic cells.

Question 78.78. Which organism would be most impacted bybiomagnification? (Points : 1.25)

eagles

rabbits

wheat plants

earthworms

corn

Question 79.79. Most of the freshwater in the world is used for(Points : 1.25)

drinking.

bathing.

irrigation.

watering lawns.

industrial needs.

Question 80.80. Which of the following is not associated withgroundwater depletion? (Points : 1.25)

subsidence

sinkholes

saltwater intrusion

lowering of the water table

sediment buildup

silverarmadillo533

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.

MICB 201 Chapter Notes - Chapter 13: Autoclave, Thermophile, Catalase

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