p.Glo Lab Report
Hypothesis:
What plates will have growth and why?: The plates with pGLO, because they are
resistant to antibiotics.
What plates will not have growth and why?: The plate with ampicillin and no pGLO,
because the bacteria are compromised by an antibiotic and do not have a recombinant plasmid.
What plates will glow under a UV light and why?: Plates with pGLO because the
plasmid contains the gene for glowing.
Background:
Transformation is the “process by which the genetic material
carried by an individual cell is altered by the incorporation of foreign
(exogenous) DNA into its genome” (MedicineNet.com, “Definition of Genetic
transformation”). Transformation in bacterial cells occurs when the cell
incorporates naked DNA into its genetic material; in a laboratory setting, this is
encouraged by placing the mixtures of transformation solution and plasmid DNA (in
+pGLO tube only) on ice, then rapidly transferring them to a hot water bath for
about fifty seconds, and then placing them back on ice again – this procedure is
called heat shock and “increases the permeability of the cell membrane to DNA”
(lab directions). The agent which the new genetic material is incorporated into
is the bacterial plasmid.
A plasmid is a circular deoxyribonucleic acid (DNA) molecule that replicates
independently of the bacterial chromosome and often is the avenue for which a
bacteria gains resistance to an antibiotic. Recombinant plasmids are those which
have DNA from two or more sources incorporated into a single plasmid. To make
recombinant plasmids, two different plasmids are cut with the same restriction
enzyme: this restriction enzyme only cuts at particular restriction sites, so the
type of cut it makes in one plasmid will be the same type of cut in another
plasmid. The cut must produce “sticky ends” so that the plasmid DNA can bind to
any other plasmid DNA with complementary base pairs. Once cut, the two plasmids
are mixed and the...