PET28c kanamycin resistance question!?
What happens if you add too high a concentration of kanamycin to pET28c plasmid, I accidently added three times as much... oops!
Asked by Brigida Bowerman 4 months ago.
It might not ruin your experiment. You may get slower growth. I haven't done this with kanamycin because most of us tend to be using ampicillin most of the time. But when someone has accidentally put twice as much ampicillin in plates, we've still wound up with colonies - we just had fewer colonies or smaller colonies or had to wait longer; I forget which. I'm sure there's some concentration at which you would simply not get growth, but I'm not sure what it would be. And since some people use 25 ug/ml and some use 50 ug/ml, I'm not sure which you're multiplying by 3. All you can do is try growing your plasmid and see what you get, or make new medium and start over from wherever you left off. I really hate pouring plates, so would probably give it a try overnight, but if I was in a hurry to get results, I'd just autoclave some new medium now and add the right amount of kanamycin. Answered by Allena Bodwin 4 months ago.
If you add more 50ug/ml of Kan E-Coli celss won't grow in media Answered by Justa Schustrich 4 months ago.
Ampicillin and kanamycin?
how does ampicillin kill bacteria and how does kanamycin kill bacteria?
Asked by Wilhelmina Chimera 4 months ago.
Ampicillin is a beta-lactam antibiotic, so while it certainly primes a bacteria for death, it is technically only bacteriostatic - preventing the bacteria from replicating. Kanamycin is an aminoglycoside. These drugs bind to the 30s subunit of bacteria ribosomes. Ribosomes are resposible for producing proteins using the cell's mRNA as a blueprint. As the protein is created, it must shift out of the ribosome to make room for the next amino acid in the chain. Aminoglycosides prevent this movement, and protein production is stalled or altered so that proteins are either no longer produced, or produced severely malformed. This is a bacteriocidal event - the bacteria dies. Answered by Noelia Alhameed 4 months ago.
Mode of action and target organisms of the following???
kanamycin, polymyxin, streptomycin, ampicillin, penicillin, bacitracin,aureomycin??
Asked by Ann Blakesley 4 months ago.
Kanamycin -disruption of protein synthesis -effective against aerobic gram-negative bacilli and some gram-positive bacteria, including mycobacteria Polymyxin -disruption of cell membrane by interacting with its phospholipids -have a bactericidal effect on Gram-negative bacilli, especially on Pseudomonas and coliform organisms Streptomycin -binds to the 16S rRNA of the bacterial ribosome, interfering with the binding of formyl-methionyl-tRNA to the 30S subunit. This prevents initiation of protein synthesis -inhibits both gram positive and gram negative bacteria, and is a therefore a useful broad spectrum antibiotic Ampicillin and penicillin -inhibits cell wall synthesis -treat urinary tract infections, otitis media, uncomplicated community-acquired pneumonia, Haemophilus influenzae, salmonellosis and Listeria meningitis. It is used with flucloxacillin in the combination antibiotic co-fluampicil for empiric treatment of cellulitis; providing cover against Group A streptococcal infection whilst the flucloxacillin acts against the Staphylococcus aureus bacterium. Of concern is the number of bacteria that become resistant to Ampicillin necessitating combination therapy or use of other antibiotics. All Pseudomonas and most strains of Klebsiella and Aerobacter are considered resistant. Bacitracin -Bacitracin interferes with the dephosphorylation of the C55-isoprenyl pyrophosphate, a molecule which carries the building blocks of the peptidoglycan bacterial cell wall outside of the inner membrane -gram positive bacteria Aureomycin -inhibits protein synthesis -broad-spectrum antibiotic Answered by Vina Berkovich 4 months ago.
What is kanamycin and what is its effects on arabidopsis thaliana?
Asked by Tawny Dronko 4 months ago.
Kanamycin is an antiobiotic. It inhibits proper function of the 30S subunit of the ribosome. Answered by Krystyna Brunton 4 months ago.
That is the scientific name for that little morsel of skin that hangs from the roof of your mouth when you bite into a really hot piece of pizza. As for the effect it has on one's Arabidopsis Thaliana (must be capitalized), that depends on how many Guiness pints you have consumed. Answered by Joline Racki 4 months ago.
Question about selectable markers - what do hygromycin, ampicillin, basta, and kanamycin do?
I know how they work for bacteria but I was asking about plants and resistance. Bacteria have a different kind of cell wall than plants but can the mechanism still be the same?
Asked by Gino Rickett 4 months ago.
Hygromycin B is an aminoglycosidic antibiotic produced by Streptomyces hygroscopicus. It is used for the selection and maintenance of prokaryotic and eukaryotic cells transfected with the hygromycin resistance gene, hph. Hygromycin B kills bacteria, fungi and higher eukaryotic cells by inhibiting protein synthesis. The resistance gene codes for a kinase (Hygromycin phosphotransferase, HPT) that inactivates Hygromycin B through phosphorylation. Cloning of the resistance gene and fusion with eukaryotic promoters has resulted in the development of vectors that permit selection for resistance to Hygromycin B in both prokaryotic and eukaryotic cells. Ampicillin is one of the most widely prescribed antibiotics. It is considered a penicillin and is a close relative of another penicillin, amoxicillin. Unlike penicillin, ampicillin and amoxicillin can penetrate and prevent the growth of certain types of bacteria, called gram-negative bacteria. Ampicillin differs from penicillin only by the presence of an amino group. The amino group helps the drug penetrate the outer membrane of gram-negative bacteria. It inhibits the third and final stage of bacterial cell wall synthesis, which ultimately leads to cell lysis. Kanamycin works by affecting 30S ribosomal subunit and causing a frame-shift or it prevents the translation of RNA. This means that instead of a codon CAT (for example in sequence CATG), a codon ATG is read by aminoacyl tRNA (aa-tRNA). Aminoacyl tRNA is consequently carrying a different amino acid, because the anticodon on the aa-tRNA is different. The protein needed cannot be synthesised - a completely different protein is synthesised or a protein similar to the one needed, but not folded correctly; it depends of the site and severness of the frame-shift. A bacterium is destroyed because it cannot produce any of its proteins correctly. Hope this info helps; I don't find anything for "basta" that looks like it relates in any way to the rest of the question.... Answered by Laura Esquivias 4 months ago.
WHAT IS THE OTHER NAME FOR KANAMYCIN INJECTION?
Asked by Jacelyn Mccleery 4 months ago.
the trade name for the the antibacterial kanamycin is Kannasyn, You could enter either in a Google search and ask for uses and side effects. Answered by Jeanelle Staten 4 months ago.
Is it possible to use kanamycin antibiotic instead of ampicillin for LB agar plate for gene expression?
I am planning to grow the colony from a host plasmid in LB agar plate. I used Kanamycin instead of ampicillin but I didn't get any colony after 16 hour incubating at 37degree centigrade temperature. What can be the possible mistake that I might have made? Any suggestion would be appreciated
Asked by Carey Leppanen 4 months ago.
The plasmid that contains the gene you want to express confers resistance to penicillins (beta-lactamase). Kanamycin is an aminoglycoside, and requires a different plasmid (one that confers kanamycin resistance. The kanamycin killed your colonies. Make new plates with ampicillin and it should work. DK Answered by Franchesca Maliszewski 4 months ago.
Why should plasmids have selectable marker(s) such as ampicillin or kanamycin? please help!?
Asked by Dorthey Mackie 4 months ago.
That way you can kill off all the bacteria that do not have antibiotic resistance The bacteria that survive after giving Ampicillin or Kanamycin contain the plasmid with the antibiotic resistance genes. In these plasmids, you can also modify them to contain a gene your interested in. That is, you can insert a gene into these plasmids. So by giving htese antibiotics, you basically are left with only the bacteria that contain these plasmids that have resistance and your gene of interest Answered by Mackenzie Marocco 4 months ago.
The selective marker is used to kill any bacteria that did not pick up the plasmid during transformation. Only bacteria that picked up the plasmid will have the gene to encode for resistance to the antibiotic used in the growth medium and selectively allows it to grow. Also, if the genes on the plasmid are not being expressed (due to not being exposed to antibiotics) the cell will kick them out. This is selective pressure. Answered by Madeline Bogar 4 months ago.
DOES LIGHT AND TEMPERATURE RENDER KANAMYCIN INEFFECTIVE?
I infused my LB agar plates with a final concentration of 30 ug/mL Kanamycin. I transformed some presumed competent E. coli with a vector containing Kanamycin resistance then spread them on one with ampicillin and one on the aforementioned kanamycin plates, and grew overnight at 37 C. the negative control didnt...
Asked by Aimee Gregory 4 months ago.
I infused my LB agar plates with a final concentration of 30 ug/mL Kanamycin. I transformed some presumed competent E. coli with a vector containing Kanamycin resistance then spread them on one with ampicillin and one on the aforementioned kanamycin plates, and grew overnight at 37 C. the negative control didnt grow but the kanamycin plate did. THE MAIN QUESTION IS, DOES LIGHT AND TEMPERATURE RENDER KANAMYCIN INEFFECTIVE? Answered by Evon Keitt 4 months ago.
Light and temperature can affect it, but people use it on bacteria plates all the time. Did you transform your competent cells with something you had just ligated or was it a DNA prep of a previously generated clone or vector... The later is supercoiled and transforms with 1000x greater efficiency than something you've just ligated together. My guess is that your transformation worked so well that you now have so many colonies on your KAN plate that they're confluent (wall to wall colonies), and the KAN is just fine. You just wound up with too many KAN resistant colonies. Storage for Kanamycin: Store at room temperature between 59 and 86 degrees F (15 to 30 degrees C) away from light and moisture. Answered by Calandra Zierden 4 months ago.
What is the specific mode of action and the target microorganism of kanamycin, polymyxin and streptomycin?
Asked by Kathey Cragg 4 months ago.
Streptomycin stops bacterial growth by damaging cell membranes and inhibiting protein synthesis. (Tuberculosis) Polymyxin works by damaging the cytoplasmic membrane of bacteria. (namely Psuedomonas, but generally gram-negative bacteria) Kanamycin affects the 30S ribosomal unit of bacteria and therefore prevents the translation of RNA. (General anti-bacterial) Answered by Myrtice Sanjose 4 months ago.