What is the gene that codes for kanamycin resistance?
All hybrid plasmids contain the promoterless part of kanamycin resistance gene (which codes for aminoglycoside 3′-phosphotransferase II) from transposon Tn5. The Km gene expression is driven by a promoters situated on pLD720.
How big is the kanamycin resistance gene?
The presumed open reading frame of the aphA-7 structural gene was 753 bp in length and encoded a protein of 251 amino acids with a calculated weight of 29,691 Da.
How does the kanamycin resistance gene work?
Resistance to kanamycin involves the activity of amino-phosphotransferase enzymes.To inactivate and modify kanamycin, an amino-phosphotransferase transfers a phosphate group from their ATP (Adenine Triphosphate) to kanamycin. Two examples of these enzymes are Aph (3′)-I and Aph (3′)-II.
What is kanamycin resistance cassette?
Taken together, we demonstrate that the kanamycin resistance gene is a suitable cassette for the genetic manipulation of T. denticola that will facilitate the characterization of virulence factors attributed to this important oral pathogen.
How do bacteria become resistant to kanamycin?
In bacteria, resistance to kanamycin is attributed to three mechanisms. One mechanism involves an aminoglycoside-modifying enzyme carried by transposons (22). The second mechanism is specific methylation of rRNA. Modification of the rRNA at position 1405 or 1408 was responsible for kanamycin resistance (5).
What is the purpose of the kanamycin resistance gene in this plasmid?
Adding an antibiotic resistance gene to the plasmid solves both problems at once – it allows a scientist to easily detect plasmid-containing bacteria when the cells are grown on selective media, and provides those bacteria with a pressure to keep your plasmid.
What is the purpose of kanamycin resistance gene?
Use in research Kanamycin is used in molecular biology as a selective agent most commonly to isolate bacteria (e.g., E. coli) which have taken up genes (e.g., of plasmids) coupled to a gene coding for kanamycin resistance (primarily Neomycin phosphotransferase II [NPT II/Neo]).
What is the difference between kanamycin A and B?
Kanamycin A has a hydroxy group at position C2′, whereas kanamycin B possesses an amino group at this position (Fig 1B). Their specific interactions with bacterial ribosomal RNAs to inhibit bacterial protein synthesis have been clearly observed through X-ray structural analysis [2].