Answer:
E.coli is a bacterium which is preffered for transformation due to its rapid growth and high efficiency of introduction of DNA molecules into cells.
Tetracycline is an antibiotic used to treat several infections such as cholera, plague, brucellosis, and malaria etcetra.
Plasmid is a small, extrachromosomal DNA molecule present in cell which is seprated from chromosomal DNA and have ability to replicate independently.
When a plasmid resistant to tetracycline and kanamycine is inserted into E.coli, the process is called transformation. E.coli might not be able to survive due to absence of plasmid but the transformed strains having antibiotic tetracycline and kanamycine will probably be able to survive it.
Hence, the culture of bacteria carrying plasmid will be able to survive only.
Maximum transformation to tetracycline achieved when cells were harvested from L broth at 5.0-6.0 cfu/ml , followed by washing twice in cold.
The susceptibility of E.coli strain to culturing transformation by plasmid DNA was examined and optimized.
Cells grown within the presence of D - cycloserine ( Cyc ) rather than Neg Gram (N x) transformed better. The presence of 5 mm Mg2+ ions in washing and CaCl2 solutions stimulated transformation about 2-fold.
Optimal conditions for transformation included a pH range of seven. 25-7.75 and a cell-to-DNA ratio of about 1.6 . 10(8) cfu/ng plasmid DNA.
The frequency of transformation was highest when cells were exposed to 100 mM CaCl2 in 250 mm.
[tex]\rm KCl + 5 mM \;MgCl2 + 5 mM \;Tris[/tex]
pH 7.6, before mixing with DNA
Therefore, the results of culturing transformed E.coli within the presence of tetracycline is justified.
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