Materials and Methods: 
      i.         The laboratory of G.P. Andrews of the University of Wyoming will provide bacterial samples.
    ii.         L. lactis both the wild type and transformed strain will be grown on M17 media that is supplemented with 0.5% glucose and incubated at 30 degrees Celsius3. We will also be streaking from frozen stalk onto TSA: Bacilis subtilis, E. coli. From frozen stalk onto blood agar plates (BPA) we will be streaking Salmonella enteridis, Yersinia enterocolitica, and Campylobacter jejuni (ADD CITATION FOR JEJUN . All will be grown in their desired 37 degrees Celsius environments.   
  iii.         L. lactis will be grown on M17 media, and in M17 broth, which will both be supplemented with 0.5% glucose and incubated at 30 degrees Celsius. We will also streak the previously stated microbes for isolation for use in later tests. We will utilize a pH meter and create an M17 broth and agar that is between the pH of 5.7 and 7.3. We will then grow the L. lactis wild type and transformed strain in the broth and measure titer using a spectometer. We will also use a similar procedure for testing the growth of L. lactis between the temperatures of 37.8 and 40 degrees Celsius. PCR will be performed using a standard PCR machine to look for plasmid presence in L. lactis after generations have been grown within the specific pH and temperature conditions3. We will also insert the naked plasmid into broth growths of the previously streaked stomach pathogens to test for the uptake of the plasmid. [RW3] We will also utilize plasmid extraction to determine the plasmids presence within the microbes (3).
 
 Data Collection and Data Analysis
Timeline:
Week 0
Streak and isolate L. lactis (wild type, and transformed), Salmonella enteridis, Yersinia enterocolitica, and Campylobacter jejuni. Make M17 broth and media, make altered pH M17 broth and media
 
Week 1
Test growth rates and abundance between the L. lactis wild type and conjugated strain. Use M17 broth, dilute down and plate to determine starting titer.
 
Week 2
Test growth rates of the L. lactis wild type and conjugated strain in the different pH, 5.7, 6.5, 7.3. Measure using dilutions.
 
Week 3
Test growth rates of the L. lactis wild type and conjugated strain in the different temperature 38.7 and 40 degrees Celsius. Measure using dilutions
 
Week 4
Determine the presence of plasmid through three generations of the L. lactis in the most successful pH. Plasmid presence will be determined by PCR.
 
Week 5
Determine the presence of plasmid through three generations of the L. lactis grown at 38.7 degrees Celsius. Plasmid presence will be determined by PCR.
 
Week 6
We will insert the plasmid into TSB broth of each of the following microbes: Salmonella enteridis, Yersinia enterocolitica, Campylobacter jejuni, B. subtilis, and E. coli  and observe if the naked plasmid is absorbed by the pathogens.
 
Week 7
Plasmid presence in the enteric pathogens will the determined by running a PCR on each.
 
Week 8
Data analysis using Microsoft Excel.
 
Frozen Stalk list 
  1. E. coli, Sept 21, 2017
  2. B. subtillus, Sept 21, 2017
  3.    
Scope: Vaccination in ungulates both domesticated and wild (cattle and elk).