THE BACTERIAL GENETICS

This lecture is the beginning of chapter 7, the doctor said that chapter 7 has 2 different topics the first one is about metabolism of the bacteria and the other topic is about the bacterial genetics. We are going to talk about genetics of bacteria and in a general idea not very deeply. An organisms genotype: it is complete collection of Bacterial Genetics genes; it is the genetics Genetics: the study of heredity. information which carries on DNA. DNA carries couple of genes it depends , if its human 46 chromosomes each one of these chromosomes in numbered 1,2,3,4, each one of these chromosomes carries number of genes , for bacteria ,the same thing, the chromosome the genome of the bacteria carries genes and it depends on the size of this microorganisms. And the numbers of genes that depends on the size of chromosomes that carry genes and gives us treats that appears on bacteria, in this case these traits can be : Pigmentation The production of certain enzyme. The production of a certain product. The type of a growth of this M.O on surface of agar Media. So we are going to call it a phenotype, An organisms phenotype refer to his physical traits such as hair, eye color in humans ,but in bacteria it is going to be: The morphology ,the type of enzyme or the type of product it is producing.

An organisms phenotype is a manifestation of the organisms genotype: whatever is carried on the gene is going to appear on appearance.
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 Genes rate all functions of the cell; In this case were going to talk about enzymes that catalyze certain reactions in the cell. Ex: polymerase is very important for DNA replication RNA polymerase is very important for transcription. Particular segments of chromosome constitutes a gene, this segment consist of couple of nucleotides, it maybe 20 of them maybe 500 of them maybe 1000 of nucleotides long. What do we mean by Mutations!

Mutations : A change in a DNA molecule we call it a genetic alteration. So lets say it ACGAAGGCCTT. If the one of these bases which make the gene changed this will lead to alteration in the genetic (myocope) Alteration it might be in this case leads to mutation , these mutation may affect on final product and it might not ,This final product might be a protein enzyme so it will not work anymore dysfunction. A change in a DNA molecule (genetic alteration) that is transmissible to offspring is called a mutation.
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 Categories of Mutations

1-Beneficial mutations: sometimes these bacteria must have a mutation as result of environmental stress .In this case it can adapt itself in a better way to the changes in the environment so in this case this kind of mutation will be beneficial mutation for the bacteria.

2-Harmful mutations (some are lethal mutations): Example: exposure to UV light For a Long Time, It Might Cause some kind of alteration or mutations in our DNA in the skin and it might leads to a cancer melanoma. The same thing for bacteria some of these mutations might be lethal to the bacteria and might kill the bacteria.

3-Silent mutations: basic maybe change in the bases but its not going to go away like there is no mutation and it doesnt effect on the Final Product.

Q: How it doesnt affect the final product which is a protein? If we look at the universal codon table well final that we have many codons give the same AA, so we might have a mutation that gives a different codon but still have the same final product the same AA.

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 Mutation rate (the rate at which mutations occur) can be increased by exposing cells to physical or chemical agents called mutagens and this Mutagens could be environmental like exposing bacteria for Radiation, certain chemicals. And this may Effect on bacterial genetic myocope and it can cause mutations, so in this case the substance that cause mutations we are going to call it mutagen. The organism containing the mutation is called a mutant and it could be: 1) wild type bacteria: for example in the researches we said the wild type bacteria its the normal bacteria, if it went through is kind of mutation then its going to become a mutant, something happened to DNA, some mutation happened in the structure of a DNA or the nucleotide sequence, base change , then it led to a new type or a different type from a wild type and were going to call it a mutant. This is the mutation How the bacteria can acquire new genetic traits ? If you exposed certain strains or species of staph aureus for example: Gram positive bacteria its wall formed by a thick peptidoglycan layer, if it is exposed to penicillin, Penecillin works on this peptidoglycan layer and it destroy it, but in this case bacteria acquired weights by which it can produce an enzyme, penicillinase: can break down penicillin .Penicillin is not anymore in the environment of this bacteria anymore. So By some kind of mutation now the bacteria are able to produce a substance can break down penicillin, this is one way, another way maybe in the old day, for example pseudomonas (gram ve bacteria) it had gates in the outer membrane in which they can introduce antibiotic that enter by these gates channel into the cell and they can kill the cell from inside. But now the bacteria one way to resist antibiotics, now we have mutations that doesnt make these gates , when these gates closed ,certain antibiotics are not going to be able to be delivered or by pass the outer membrane to get inside the cell, so its not going to be affected against pseudomonas .And we know that
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staph aureus and pseudomonas both of these a problem in the hospital, they are nosocomial infection. The problem of them that we cant use antibiotic that against them because those type of bacteria they developed new ways to resist bacteria ( which have mutations ) so also in this case we have to develop new kinds of drugs or treatment to kill this M.O or they are going to kill us . So how they can acquire genetic traits? For example Bacteria have resistance for antibiotics and other doesnt have it. But the one that has resistance for antibiotic can give its gene to the one doesnt have it so now both of them have antibiotic Resistance to a certain antibiotic drug. How they can exchange genetic material?so we have something named: 1-lysogenic conversion 2-transduction 3-transformation 4-conjngation An extra chromosomal DNA molecule is called a plasmid. An organism that acquires a plasmid acquires new genes. A plasmid can exist by itself or can integrate into the chromosome is called an episome. Its not really a part of chromosomal DNA. In bacteria the chromosomal DNA which carries the genotype of the bacteria that is needed for metabolism and structure of the cell replication ..etc .there is also extra chromosomal DNA found in the cytoplasm. It is smaller than the chromosomal DNA, which means the numbers of genes that are carried on the plasmid are less than what carries on the chromosomes.

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We have families of plasmid they have members, for example there are important plasmid for antibiotic resistance also we have important plasmid for metabolism, that makes a specific A.A. So we have different kinds of plasmid. Note that ! Plasmid can either exist by itself or it can integrate itself into the chromosome, we call it an episome.

Here the doctor showed a picture:

The left picture illustrates a plasmid that is expelled outside the E.coli bacteria. The right picture illustrates a closer view of the plasmid, which can carry essential genes

1- Lysogenic conversion:
There is something called bacteriophage; which is a virus that can infect the bacteria. We have a type of bacteriophage that is called temprate phages. These are considered a lysogenic phages; which means they can inject their DNA into the bacterial cell and that DNA will be integrated into the bacterial chromosome.

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It will not cause a lytic cycle, so we will have a bacteria that have the bacteriophage DNA. That DNA may lay down there for a long time until the environment inside and outside the cell will be improved, and then they may excise themselves from the chromosome, taking genes from the bacterial chromosome. They will replicate inside the cell, synthesizing more and more DNA containing the original bacteriophage DNA and some part of the bacterial DNA and protein that will form the virus inside the bacteria. They will form holes in the membranes of the bacteria cell and will come out of the bacteria. And will attack another nearby bacteria by this it will transfer genes from one bacteria to another. So this is another way which they can transfer genes from one species to another. These viruses if came from bacterial cell to another then they will carry some of the genes from previous species to the next one and then we have lysogenic conversion A Phage is called prophage when it consists of the DNA without the virus coat there is no virus coat only the DNA. This prophage will integrate itself in the bacterial chromosome, as mentioned previously, the bacteria that contains the prophage is called lysogenic bacteria.

2-Transduction to carry a cross:

Also involves bacteriophage. In transduction the bacteria genetic material came across from one bacteria cell to another by a bacterial virus bacterium acquire new bacterial genes. All the small amounts of genetic material are transferred by transduction which is going to take, once it going to be excised from the chromosome, It will take simple parts from bacteria DNA, from left and right which surround the Virus DNA. From both slides, it takes simple parts from bacteria DNA, from left and right which surround the virus DNA.
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Note how this differs from lysogenic conversion; where in bacteria acquire new genetic information in the form of viral genes.

3-Transformation:
We have 2 types of transformation:1) Transformation by nature: it can happen by nature. 2) Transformation by force: that we use it in researches of microbiology etc were trying to introduce by force certain genes into the bacterial cell. In the transformation: a bacterial cell becomes genetically transformed following the uptake DNA fragments or naked DNA. In transformation we will have a bacteria and a naked DNA floating around the cell. This bacteria must be competent cell; that means it has all the chromosomes and machinery that is required to take the nearby virus. This naked DNA will be introduced in the chromosomal DNA of the bacteria, and new characteristics will be acquired in that bacteria. All the small amounts of genetic material are transferred by excise from the chromosome. The ability to absorb naked DNA into the cell called competence and the bacteria which has the ability to absorb DNA is called competent bacteria. Not all species of bacteria are able to take in any kind of naked DNA around it has to be competent , it has to be able to open up cell may to introduce this named DNA. for example, in molecular microbiology researches purposes we make the cell competent by using different chemicals and we force certain DNA to get into the cell and so introducing new traits that we are controlling into this bacterial cell it will be under control and there for we can make a lot of researches , Ex: certain

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genes, what theyre producing . Etc a lot of things have been discovered by that. For example in microbiological researches for..purposes and etc we make the cells competent by using different chemicals and techniques we make it competent and we force certain DNA to get into the cell and by this we are introducing new traits that we are controlling into this bacterial cell and depending on this we can do a lot of studies and a lot of things had been discovered as a result of transformation. Transformation properly is not widely in nature because not all the species that you can find are competent to take naked DNA.

4-Conjugation:
The other way by which bacteria can acquire new traits, new genetic and chromosomes: Conjugation. It Is a widely used way, it is a phenomena in the bacteria and we manipulated this procedure tonature to our own benefits on molecular microbiology laboratory tubes. Conjugation: means mating between 2 bacteria. In this case it involves specialized type of pillus called a sex pilus we have two types of pillus: sex pillus and attachment pillus Bacteria that have the sex pillus: donor cell Attaches by a means of the sex pilus to another Bacterial cell called the recipient cell. The donor cell is going to extend its sex pillus to the recipient cell. In this case they have close contact they have to be in close, close to each other. Structure of this pillus is tubes and the bacteria that have got the Pilli also contain plasmid. The plasmid carries certain genes on it. It Does replication and give a
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copy in the donor cell and another copy that pass by the Pilli to the recipient cell. And at the end of this both of these they are going to have the same plasmid. So the donor cell and the recipient cell have the plasmid and both plasmid carry certain genes of them depends on the type of the plasmid. If we have R plasmid which is one of plasmid types: resistance plasmid which can carry genes which can resist certain antibiotics. It might be penicillinase, tetracyclonase. Plasmid that contains multiple genes for antibiotic resistance IS known as a resistance factor or R-factor. The bacterial cell that receives an r factor becomes a super bulk it is not going to care if this antibiotic surrounding it or not, because it has the gene that can break down this antibiotic and that what happens in the pseudomonas bacteria, that causes a nosocomial infection.

Here the doctor started to talk about nosocomial infection: Nosocomial infection is the infection that is acquired while the patient is in the hospital. There are a lot of bacteria responsible for it: 1) Pseudomonas bacteria. 2) Acinitobacter this bacteria has acquired a resistance for multiple antibiotics and that can cause the death of the patient whom infected by it 3) Klebsiella 4) Staph aureus, esp. the MRSA type MRSA: methicillin resistance staph aureus. All of these bacteria have resistance for multiple antibiotics, esp. the pseudomonas and staph aureus. So if an immunocomprimised patient got infected by these microorganisms, he might die.

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You have a picture in the slides that show the conjugation process of the E.coli bacteria, and it illustrates the sex, and attachment Pilli.

Genetic engineering:
Genetic engineering or recombinant DNA technology involves techniques to transfer eukaryotic genes (particularly human genes) into easily cultured cells to manufacture important gene products (mostly proteins). A recombinant gene has been used in different fields: 1) Production of vaccines, like hepatitis b, tetanus: Lets take how the recombinant gene technology has been utilized to produce the vaccine against hepatitis b virus: This vaccine is composed of the surface antigen of hepatitis b virus, not the whole virus, (it is a subunit vaccine).once introduced to the body, our body will produce antibodies against this surface antigen, these antibodies will remain in our body to prevent the infection of hepatitis b virus.

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How did we make that surface antigen of hepatitis b virus? We took the gene that encodes the hepatitis b surface antigen from the hepatitis b virus, and then we put it in the vaccine virus. Vaccina virus is not harmful. So we have vaccina virus that has hepatitis b surface antigen. Hepatitis b virus vaccines are taken in 3 booster doses. A note from a question that has been asked in the lecture: vaccines can rarely cause a disease in the human. 2) Production of insulin to treat DM: Plasmids are frequently used as vehicles for inserting genes into cells: In labs we can manipulate the plasmids and inject a certain gene inside it by a process that requires a lot of enzymatic action, (lets suppose that gene is the gene responsible for the production of insulin.) Then we let the E.coli bacteria takes that plasmid in the process of transformation, now we have an E.coli bacteria that has the plasmid containing the insulin gene inside it. Now we let the bacteria multiply and produce the insulin. We take the bacteria and remove the cell wall and take the insulin. Buy this we have produced a lot of amount of insulin 3) Production of clotting factor to treat hemophilia, production of growth hormones. 4) Antibiotics: many antibiotics that are found in the markets are produced by using the streptomycin bacteria. 5) Synthesis of antibodies: some antibodies are important to neutralize certain toxins, other antibodies are used in research; for example the antibodies can be tagged by florescent or radioactive material, in this case we can identify the antigen that the antibody will bind to.

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Gene therapy:
Gene therapy of human diseases involves the insertion of a normal gene into cells to correct a specific genetic disorder caused by a defective gene. A cell can have an abnormal gene, which causes a disease, here we can inject the normal gene into a virus and then we inject that virus in the body. It will go to the cell and then there will be an exchange of the gene material. The cell will take the normal gene. And by this we can cure the disease. Not all viruses can be used to do that, for example we can use adeno viruses or vaccina viruses. Genes may someday be regularly prescribed as drugs in the treatment of diseases (e.g., autoimmune diseases, sickle cell anemia, cancer, cystic fibrosis, heart disease, etc.)

This Script is Done by:

Jumana Ali Al-shawabke. Rana Al-Otoom.

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