Question

1)

describe the results of the transformation experiment (E.coli)

2) identify one ethical concern are raised by CRISPR experiment (E.coli)

Answer 1

Results of transformation experiment of E.Coli are follows:-

1.E coli can be rendered competent for DNA uptake by a temperature shock(0 degree celcius to 42 degree celcius and then to 0 degree celcius) of the recipient cells.

2.Frequency of transformants was enhanced by a factor of 10 when the transformation mixture was plated on selective Tris-based minimal salts medium containing 10 mM Ca2+ and 10 mM Mg2+ instead of on conventional phosphate - based minimal medium.

3.The DNase treatment if applied before the heat shock was sufficient to reduce the transformation frequency to less than 10 raised to the power -7 transformants per recipient cell.

4.The high transformation frequencies obtained when DNA was added after the heat shock,it clearly show that cells were rendered competent for DNA uptake by a heat shock in the absence of DNA.

5.Shortly after the temperature shock (from 0 degree celcius to 42 degree celcius) increase in cell uptake ability is mainly due to increase permeability of outer membrane,but after 1 hour further incubation (at 0 degree celcius) the total increase in beta lactamase(Found in periplasmic space located between the cytoplasmic membrane and the outer membrane )activity was due to enzyme that had leaked out of periplasm.

Ethical concerns of CRISPR(Clustered regularly interspaced short palindromic repeats) experiments of E.coli are follows:-

1.The extent to which CRISPR use should be permitted.

2.Access to CRISPR applications .

3.Issues related to genome editing.

4.Wheter international regulations governing inappropriate CRISPR utilization should be crafted and publicized.