Kanamycin resistance

Dideoxykanamycin B active against kanamycin-resistant Escherichia coli and Pseudomonas aeruginosa," H. Umezawa, S. Umezawa, T. Tsuchiya, and Y. Okazaki, J. Antibiol., 24 (1971) 485-487. 

Tc = Tetracycline resistant Km = Kanamycine resistant Ap = Ampicilin resistant... 

A) Organization of the papl-D allele and the structure of pMN-PAP1. Bastar, Basta resistance Kafr, kanamycin resistance LB, left border pUC19, pUC 19 plasmid RB, right border 4 x 35S denotes four copies of 35Se. 

Mob, can be mobilized during conjugation with broad-host-range plasmids Xm, kanamycin resistance Km, kanamycin sensitivity Cm, chloramphenicol Sm, streptomycin 7c, tetracycline. 

Clones with ampicillin resistance but without kanamycin resistance are examined with the CloneChecker System. By this antibiotic selection, the undesired vector heterodimer is removed. If both of the clones are false, additional clones are analyzed using the same protocol. When appropriate clones are obtained for almost all of the genes, they are inoculated from the ampicillin plates to prepare plasmids in a 96-well format by using the Wizard SV 96 Plasmid DNA Purification System see Note 6). 

Strains of multidrug-resistant Mycobacterium tuberculosis, including streptomycin-resistant strains, are usually susceptible to amikacin. Kanamycin-resistant strains may be cross-resistant to amikacin. The dosage of amikacin for tuberculosis is... 

These kanamycin-resistant plants contain the foreign gene. 

In 1993, suicide by an E. colt containing the npt -sacFL-B suicide cassette (nptll gene encodes kanamycin resistance) was reported (Recorbet et al., 1993). In the absence of sucrose the sacK gene does not induce expression of the sacB gene. To induce cell death, sucrose was added to the soil in which the organism was released. This causes the cells to produce levansucrase (a sacB product), which in turn causes the periplasm of the cells to fill with levan. This leads to cell death by lysis. Sucrose... 

Once the desired plant variety has been selected, the marker gene serves no useful purpose in the new plant, but it does continue to produce the gene product, APH(.V)I1 in the case of kanamycin resistance. This enzy me is present at very low concentrations in food. 

The use of marker genes that encode resistance lo clinically important antibiotics raises questions regarding whether the enzyme in the foot could inactivate oral doses of the antibiotic or whether die gene present in the plant DNA could be transferred lo pathogenic microbes in the GI tract or in soil rendering them resistant to treatment with the antibiotic, FDA evaluated these questions for the use of kanamycin resistance in tomato, cotton, canola,... 

There is no known mechanism by which a gene can be transferred from a plant chromosome to a microbe. Thus, the possibility of that such transfer would generate new resistant organisms is very small, especially when compared to the high rate of spread of resistance through known mechanisms of microbe to microbe transfer to antibiotic resistance. FDA believes that the use of marker genes that encode resistance 10 other clinically useful antibiotics can be evaluated by similar criteria that were used for kanamycin resistance. 

Fig. 7.11 Schematic diagram of foreign gene expression in plant. P promoter, T terminator, pGA482 plant cloning vector, Km kanamycin resistance gene, Tc tetracycline resistance gene.

The His-tagged MBP-MS2 coat fusion protein (HMM) used in this method was created to allow the protein to be immobilized on both Ni2+ or amylose affinity resins. HMM is a 59-kDa protein containing an NT-terminal hexahistidine (6x His) tag, a central maltose-binding protein (MBP) domain, and a C-terminal MS2 coat protein containing the V29/dIFG mutations, which prevent protein multimerization and increase its affinity for RNA (Lim and Peabody, 1994). The protein is expressed in E. coli from plasmid pHMM, which confers kanamycin resistance, and purified using affinity chromatography. 

ImmEI—immune to colicin El, Ap —ampicillin-resistant, CmR—chloramphenicol resistance, Km —kanamycin-resistant, Tc —tetracycline-resistant. 

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