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  • During mismatch repair, how does the enzyme recognize which is the new and which is the old strand?
  • How does an intercalating agent introduce a mutation?
  • What type of mutation does photolyase repair?

Identifying bacterial mutants

One common technique used to identify bacterial mutants is called replica plating . This technique is used to detect nutritional mutants, called auxotroph s, which have a mutation in a gene encoding an enzyme in the biosynthesis pathway of a specific nutrient, such as an amino acid. As a result, whereas wild-type cells retain the ability to grow normally on a medium lacking the specific nutrient, auxotrophs are unable to grow on such a medium. During replica plating ( [link] ), a population of bacterial cells is mutagenized and then plated as individual cells on a complex nutritionally complete plate and allowed to grow into colonies. Cells from these colonies are removed from this master plate, often using sterile velvet. This velvet, containing cells, is then pressed in the same orientation onto plates of various media. At least one plate should also be nutritionally complete to ensure that cells are being properly transferred between the plates. The other plates lack specific nutrients, allowing the researcher to discover various auxotrophic mutants unable to produce specific nutrients. Cells from the corresponding colony on the nutritionally complete plate can be used to recover the mutant for further study.

  • Why are cells plated on a nutritionally complete plate in addition to nutrient-deficient plates when looking for a mutant?

The ames test

The Ames test , developed by Bruce Ames (1928–) in the 1970s, is a method that uses bacteria for rapid, inexpensive screening of the carcinogenic potential of new chemical compounds. The test measures the mutation rate associated with exposure to the compound, which, if elevated, may indicate that exposure to this compound is associated with greater cancer risk. The Ames test uses as the test organism a strain of Salmonella typhimurium that is a histidine auxotroph, unable to synthesize its own histidine because of a mutation in an essential gene required for its synthesis. After exposure to a potential mutagen, these bacteria are plated onto a medium lacking histidine, and the number of mutants regaining the ability to synthesize histidine is recorded and compared with the number of such mutants that arise in the absence of the potential mutagen ( [link] ). Chemicals that are more mutagenic will bring about more mutants with restored histidine synthesis in the Ames test. Because many chemicals are not directly mutagenic but are metabolized to mutagenic forms by liver enzymes, rat liver extract is commonly included at the start of this experiment to mimic liver metabolism. After the Ames test is conducted, compounds identified as mutagenic are further tested for their potential carcinogenic properties by using other models, including animal models like mice and rats.

Diagram of the process of identifying auxotrophic mutants. First a medium containing histidine grows colonies of bacteria. Then, a sterile velvet surface is pressed onto the plate to pick up cells from bacterial colonies. Next the cells on the velvet are transferred to new plates, one with histidine and one without. A mark on the plate ensures that the orientation of the colonies is the same for all plates. After the plates are incubated, compare the growth on the plates to identify auxotrophic mutants that grow on medium containing histidine but do not grow on medium lacking histidine. A colony that is missing on the medium lacking histamine but present on the medium with histamine is the auxotrophic mutant.
Identification of auxotrophic mutants, like histidine auxotrophs, is done using replica plating. After mutagenesis, colonies that grow on nutritionally complete medium but not on medium lacking histidine are identified as histidine auxotrophs.

Questions & Answers

can you please differentiate between vacuoles and inclusion bodies
Noor Reply
vacuoles consists of watery solution called cell sap while bodies are organelles
Prince
5 difference please
Noor
Archaea is an odd type of bacteria that live in extreme environments; high salt, high temperature etc. usually called extremophiles.
Danesia Reply
you know anyone that has delt with enterobacter agglomerans.
Vernon
it is a type of gram negative bacteria and its ubiquitous mostly isolated from plant stuffs and also from faecal material of animals. I heard the name pantoea . it is bacillus in structure.
Sujan
What are the classification of microorganisms
Portia Reply
Basic classification are eukaryotic and prokaryotic..
micro organisms are classified into three namely: Archaea, eukaryote and prokaryotic organisms
Blandine
Yay that's right
what is archaea
Ateeq
I've got an issue with the message view
Frank Reply
Is there any effective way to learn microbiology.
Frank Reply
I have some basic knowledge of biology. I mean i'm totally beginner in that field.
Frank
Assuming that i'm interested in neuroscience. What is the best way to master that particular subject.
Frank
Hi
Can you wait me just a minute
Iam apologies for guidance you in neuroscience.. I'm not physician.. I'm microbiologist
My pleasure to join to you
Let's begin a conversation
Frank Reply
What would be saying
?
Akram
do you know anything about microbiology
diana
I have so much studying to catch up on🙄
Britnee
any study tips
Britnee
Yas of course
What would you like to know
well i'm studying Pharmacy second year and i just started With Microbiology
diana
if you have any basics to tell us 🤷🏻‍♀️ would be helpful
diana
Yas, first you must know that you will have a very easy lecture..
In second stage you will try to learn the type of microbiology
A simple definition for microbiology
In second stage you will try to learn the type of microbiology.. you must learn the bacteria..its structure.. Classification of bacteria according to shape..
It's so easy
What triggers the natural killer cells
jenipher Reply
microbiology is the study of science which deals with study of microorganism e.g .virus,bacteria
sufian Reply
microbiology is a branch of science that deals with invisible microorganisms.
Abdinur
microbiology is the branch of science which deals with the study of organisms too small to be seen with the naked eye
Andrre
what is microbiology
Safwan Reply
history of microbiology
julie
give all details about cell
Samim Reply
Give me another names for all hypersensitivity
Don Reply
Give me the modern father of microbiology
Don
Antonie van Leeuwenhoek
RITU
Louis pasteur is father of modern microbiology Antonie van Leeuwenhoek is only the father of microbiology
Sujan
Antonie van Leeuwenhoek was the first person to observe the mos. whereas pasteur is the one who described about the vaccine , pasteurization etc.
Sujan
what is the virulence factor
mukhtar Reply
is the abilityof pathogen that cause disease
mary
classification of bacteria
Divya Reply
are classified into 5 as follow 1= bacilli 2= vibro 3= spirilla 4= cocci 5= spirochaetes.
Abdulkareem
this one is on the basis of morphological structure
Sujan
gram positive and gram negative according to gram staining
Sujan
u can find on the basis of temperature resistance and oxygen tolerance
Sujan
good
ANAS
on basis of oxygen requirement they could be categorized as Aerobic anaerobic facultative aerobes microaerophilic
ANAS
bacteria are classified into the three type according to their shape 1.Round cocci 2.Rod bacilli 3. spiral
mukhtar
sorry to say but for microbes there is no facultative aerobes instead it must b facultative anaerobic and obligatory anaerobic, aerotolerant
Sujan
Which ones are the microaerophilic
Godwin
what is the difference between krebs cycle and calvin cycle
Baye Reply

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Source:  OpenStax, Microbiology. OpenStax CNX. Nov 01, 2016 Download for free at http://cnx.org/content/col12087/1.4
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