Gene Transfer Lab Outline Essay

712 Words Feb 4th, 2012 3 Pages
I. TITLE: GENE TRANSFER OF ANTIBIOTIC RESISTANCE AND BIOLUMINESCENCE IN ESCHERICHIA COLI

II. INTRODUCTION

A. Background on antibiotics

1. History of antibiotics

a. Fleming’s discovery of penicillin and its early significance

-antibiotic inhibition of protein synthesis, cell wall synthesis, and other cellular processes

b. Antibiotic overuse led to bacterial resistance

Background on the transfer of resistance genes from one bacterium to another

1. Location of genes in bacteria and their means of transfer

a. Genes located on plasmids, “mini-chromosomes” in bacteria and carry few traits

b. Direct transfer by transformation of gene transfer from one bacterial cell to another (Purpose)

C.
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10 micro-liters of plasmid DNA pipetted into test tube #2 and both incubated for 20 minutes

3. Both test tubes transferred to water bath for 2 minutes (heat shock)

4. Immediately placed on ice for 5 minutes

5. 900 micro-liters of LB medium added to both test tubes (no ampicillin yet) and incubated at 37 degrees Celsius for 60 minutes

a. allows recovery of cells before exposing to antibiotic

B. Preparation of petri dishes

1. Labeled 4 petri dishes A, B, C, & D

a. Plates A & B contain LB agar

b. Plates C & D contain LB agar plus ampicillin

c. 100 micro-liters from tube #1 onto plates A & C

d. 100 micro-liters from test tube #2 onto plates B & C

2. Separate sterile glass bead placed in each petri dish

a. Rolls around and spread cells equally throughout

3. All plates incubated for 12 hours in incubator at 37 degrees Celsius then placed in refrigerator for one week

4. All plates were warmed to active bacteria for assessment of bioluminescence

RESULTS

TABLE 2: ACTUAL RESULTS FOR PLATES

Plate with description |Colonies present

(yes or no)

(if so, size of colony) |Plates capable of bioluminescence | |Plate A (no plasmid DNA or ampicillin) from test tube #1 |Yes, lawn plate (too many to count). Dense growth. |No | |Plate B (+ plasmid DNA, no ampicillin) from test tube #2 |Yes, lawn plate (too many to count). Dense growth. |No** | |Plate C (no plasmid

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