Pare Project

Jaqueline Jacuinde
Vanessa Ortiz
Candy Hernandez Fraga
Solomon Larsen

Fall 2022
Jo and Hannah
 Introduction

The purpose of this experiment is to learn about the the

possible antibiotic resistant bacteria found in our
environment. This project allowed us to learn more about
potential antibiotic resistance genes that exist in nature.
For example we completed a series of tests on dirt from the
countryside of Las Vegas in order to possibly identify
bacteria with antibiotic resistant DNA.
 Soil sample collection site
Information about soil collection site

Date of collection 11/1/2022

Countryside (not heavily populated)

No domesticated animals visit frequently

Nearest body of water Lake mead (approximately

25 miles away)

Sample was not collected on a farm (no livestock

nearby)
 Pare collection Data Results
For our chosen agar plates we choose
colonies from out PS1 with 3tet and
PS2 NA.

Most of our agar plates showed very

little colony growth. This could have
been due to plating mistake or due to
incubation at a wrong temperature.
Pare Collection
Data
 Agarose gel Electrophoresis results
The purpose of the Agarose gel electrophoresis is to separate
DNA fragments following restriction endonuclease digestion or
PCR amplification ( the DNA is cut by enzymes and fragments
are replicated )

We are Interested in Finding the 16S which is 1450 bp.

We did cells 3 and 4. We can only see 1 line in cell 3 around the
3000bp mark.

The single line in cell 3 at around 3000 bp could mean 2 thing.

1. The DNA sample came from a part of the cell that didn't have
the portion of DNA we were interested in

2. The restriction enzyme wasn’t given enough time to fully

digest the DNA sample before it was loaded into the gel.

The absence of a line in cell 4 most likely means we got the

DNA from a membrane protein or some other specific part of
the cell it could also mean we didn't add enough of the DNA
sample to the cell
 DNA sequencing results
>SampleA Caulobacter sp (A-Proteobacteria)
NNNCNNNNNNNNCNNNCNNCAAGGNACCTGACTCCTTGCGGATAGCGCAGCGCCTTCGGGCAAGGCCAACTCCCATGGTGTGACGGGCGGTGTGTACAAGGCCAGGGAACGTATTCACCGCGGCATGCTGATC
CGCGATTACTAGCGATTCCAACTTCATGCTCTCGAGTTGCAGAGAACAATCCGAACTGAGACGACTTTTAGGGATTGGCTCCCCCTCGCGGGATTGCAGCCCTCTGTAGTCGCCATTGTAGCACGTGTGTAGCCCA
CCTTGTAAGGGCCATGAGGACTTGACGTCATCCCCGCCTTCCTCCGGATTAACTCCGGCAGTACGATTAGAGTGCCCAGCCAAACCTGATGGCAACTAATCGCGAGGGTTGCGCTCGTTGCGGGACTTAACCCAAC
ATCTCACGACACGAGCTGACGACAGCCATGCAGCACCTGTGTCCCAGTCCCCGAAGGGAAAACCACATCTCTGTGGCGGTCCAGGCATGTCAAAAGGTGGTAAGGTTCTGCGCGTTGCTTCGAATTAAACCACAT
GCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTTAATCTTGCGACCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCGTCACCGACATGCATGCATGCCGACAACTAGCACTCATCGTTTACG
GCGTGGACTACCAGGGTATCTAATCCTGTTTGCTCCCCACGCTTTCGAGCCTCAGCGTCAGTAACGGACCAGTATGTCGCCTTCGCCACTGGTGTTCTTCCGAATATCTACGAATTTCACCTCTACACTCGGAGTTCC
ACATACCTCTTCCGTACTCAAGACTGCCAGTATCAAAGGCAATTCCAAGGTTGAGCCCTGGGCTTTCACCTCTAACTTAACAGTCGGCTACGCTCCTTTACCCCAGTATTCCGAGCAACGCTAGCCCCTTGTATTACG
CGGCTNTGGGNCGAAGTAGCGGGGCTCTTTCCGGTACGTATTACNGCCCGGGAAAAAATTACATCCTAAACTTATCATCCGCGAAGGCTGGGTAGTT
>SampleB Massilia Kyonggiensis (B-proteobacteria)
NNNNNNATCTACCGTGGTAGCGCCCTCCTTGCGGTTAAGCTACCTACTTCTGGTAAAACCCGCTCCCATGGTGTGACGGGCGGTGTGTACAAGACCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACT
AGCGATTCCAACTTCACGCAGTCGAGTTGCAGACTGCGATCCGGACTACGATACACTTTCTGGGATTAGCTCCCCCTCGCGGGTTGGCGGCCCTCTGTATGTACCATTGTATGACGTGTGAAGCCCTACCCATAAGG
GCCATGAGGACTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCATTAGAGTGCTCTTGCGTAGCAACTAATGACAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCT
GACGACAGCCATGCAGCACCTGTGTTCAGGTTCCCTTTCGGGCACCCTCCAATCTCTCGGAGGTTCCTGACATGTCAAGGGTAGGTAAGGTTTTTCGCGTTGCATCGAATTAATCCACATCATCCACCGCTTGTGCG
GGTCCCCGTCAATTCCTTTGAGTTTTAATCTTGCGACCGTACTCCCCAGGCGGTCTACTTCACGCGTTAGCTGCGTTACCAAGTTAATTAAAACCCGACAACTAGTAGACATCGTTTAGGGCGTGGACTACCAGGGTA
TCTAATCCTGTTTGCTCCCCACGCTTTCGTGCATGAGCGTCAATCTTGACCCAGGGGGCTGCCTTCGCCATCGGTGTTCCTCCACATCTCTACGCATTTCACTGCTACACGTGGAATTCTACCCCCCTCTGCCAGATT
CAAGCCTTACAGTCTCCATCGCAATTCCCAGGTTGAGCCCGGGGCTTTCACGACAGACTTACAAAACCGCCTGCGCACGCTTACGCCCAGTAATTCCGATTACGCTTGCACCCTACGTATTACCGCGCTGCTGGGN
CGTNGTTAGCCGGTGCTTATCTTCAGGACGTCATAGCCCAGAAATAGCCTGAACGTTCTTCCTGACAAANAGCTTTAAACCGAAGGCTTCTCATCCGCGCATGGGGATAAGCTGCGCCTTGCCAAATTCCCCGGCG
CTCCGANNNNNTGGACGGTTCATTCAGGGNNGGCCCCNCNNNAGGTTTGANNNCCCTGGGAACTTACCNNCCATAAAAATATCCCCAAGAAAAGTTTGGCCCGTTTTTNNNANANNAGGATTTATTTTCTNCCCCTN
NGGTANCATTCCNNNCCCCCCG
Blast results and Identification
Our 2 samples gave us 2 different bacteria.

The first was a 96% match for Caulobacter sp (Genus Caulobacter) or A-Proteobacteria (Class). This is
a Gram-Negative Bacteria Normally found in freshwater lakes and streams. And rarely cause disease

The 2nd sample was 86% match for Massilia Kyonggiensis (Genus Massilia) or B-Proteobacteria
(Class). It is a gram negative rod shaped bacterium.
Best Alignment

>AA_18099 (Caulobacter sp)

>AA_18100 (Massilla)
 Summary of Results

After exposing the bacteria found in our dirt sample to antibiotics , we discovered Caulobacter
sp and Massilia Kyonggiensis . These bacteria’s harbor resistant genes to antibiotics. These
finding can be useful for us to have a better understanding of how soil may be exposed to high
levels of antibiotics which then cause the presence of more antibiotic resistant microbes in the
environment , which can be linked to clinical infections.