ABSTRACT

The Illinois Junior Academy of Science

CATEGORY

Microbiology

STATE REGION #

SCHOOL

Niles North High School

IJAS SCHOOL #

6038

CITY/ZIP

Skokie, IL 60077

SCHOOL PHONE #

847.626.2254

SPONSOR

Christi Camel

MARK ONE:

EXPERIMENTAL INVESTIGATION X

NAME OF SCIENTIST*

Med Jimenez

DESIGN INVESTIGATION
GRADE

NAME OF SCIENTIST

GRADE

NAME OF SCIENTIST

GRADE

NAME OF SCIENTIST

GRADE

12

* If this project is awarded a monetary prize, the check will be written in this scientist's name, and it will be his/her responsibility to distribute
the prize money equally among all participating scientists.

PROJECT TITLE

Antibiotic Resistance of Escherichia coli on Rifampicin caused by Mutations within the

Purpose: Antibiotic Resistance is created towards rifampicin resistance with a 4-methyl-1-piperazinaminyl group using

DH5 Escherichia coli strain. This is used to determine how the rifampicin inhibited the bacteria from performing binary
fission and eventually causing it to undergo cell apoptosis. Transcription factors are also looked with the bacterial RNA
Polymerase and analyzed using a Polymerase Chain Reaction and a sequencing and genotyping facility at the University of
Chicago. The rpo gene will be used to determine the allosteric sites of inhibition causing the mutations within the beta
subunit pocket groove. After sequencing the gene, the mutated DNA from the rifampicin resistant bacteria will be
compared to the wild type DH5 Escherichia coli strain and analyzed for points of mutations using Sequencher.
Procedure: The wild type, DH5 Escherichia coli, is first exposed to various levels of Rifampicin and the mutant strains that evolve
around the rpo gene and allow RNA Polymerase and transcription to occur, will be selected, and put through a

thermocycler in order to perform Polymerase Chain Reaction. The finished product created by using rpo1240F and
rpo2226R primers will undergo gel electrophoresis and and ExoSAP procedure before being sent out to The University of
Chicago sequencing and genomic facility to be sequenced. After getting the wild type and the mutated DH5 Escherichia
coli strains, the protein will be modeled using Sequencher and analyzed for steric hindrance, rotamers, and hydrogen
bonding to determine the type of interaction and what type of mutation occurred in the DH5 Escherichia coli strain and
the effects of the rifampicin with a 4-methyl-1-piperazinaminyl group.
Conclusion: In the end of the experiment, it can be concluded that the antibiotic resistance of DH5 Escherichia coli on the

Rifampicin is caused by a single point mutation. Because of this mutation, the transcription factors and the RNA
Polymerase is able to continue on because it the rifampicin no longer fits in the subunit groove of the bacterial DNA.
Further conclusions from this in vitro method shows that the preserved DNA region remains true, the mutation rate cannot
be high around this area as the RNA Polymerase is an essential enzyme for the bacteria. Also, reproductive stress is evident
from the results gathered from the first part in that even though the bacteria survived, the rate of reproduction seemed to
have slowed down in the environment.
1) Limit Abstract to 3 paragraphs (about 200 words or less). a) Purpose - what you set out to investigate; b) Procedure - how you did
it; c) Conclusion - based on your results. Label each paragraph.
2) Must be typed, single-spaced on the front of this form. Do not write on the back of this form.
3) Three copies of your complete paper are required at the State Science Project Exposition.
Four copies of your complete paper are required for the State Paper Session Competition.
This form must be used. This form must be displayed on the front of the exhibitors display board. It may be reduced to half a sheet of paper; 8.5
inches (vertical) X 5.5 inches (horizontal).

SAFETY SHEET
The Illinois Junior Academy of Science

Directions: The student is asked to read these introductions carefully and fill out the bottom of this sheet. The
science teacher and/or advisor must sign in the indicated space. By signing this sheet, the sponsor
assumes all responsibilities related to this project.
Safety and the Student: Experimentation or design may involve an element of risk or injury to the student, test
subjects and to others. Recognition of such hazards and provision for adequate control measures are joint
responsibilities of the student and the sponsor. Some of the more common risks encountered in research are
those of electrical shock, infection from pathogenic organisms, uncontrolled reactions of incompatible
chemicals, eye injury from materials or procedures, and fire in apparatus or work area. Countering these
hazards and others with suitable safety practices is an integral part of good scientific research. In the chart below,
list the principal hazards associated with your project, if any, and what specific precautions
you have used as safeguards. Be sure to read the entire section in the Policy and Procedure Manual of the Illinois
Junior Academy of Science entitled "Safety Guidelines for Experimentation" before completing this form.
Possible hazards

Precautions taken to deal with each hazard

-Acids and Stains, handled in a bio safety lab and used according to the instructions and materials handout in the lab.
After use, they have been properly disposed of.
-Cell cultures are also handled in this way. All were autoclaved and bleached before being thrown away. Careful and
meticulous techniques were used the whole time and the cells were handled with care
-Glass objects were also handled with care using sterile and proper techniques while using the material.
-Lab coat, googles, and gloves were worn during the whole procedure of the lab
Specific safety practices related to materials requiring endorsement sheets should be detailed on the specific
endorsement sheet and not included on this safety sheet.
Please check off any other possible endorsements needed. Include these documents in your paper and on your board.
_____Humans as Test Subjects for any projects involving humans including survey administration;
X Microorganisms-for any projects involving bacteria, viruses, yeasts, fungi or protazoa;
_____Non-Human Vertebrates -for any projects involving fish, amphibians, reptiles, birds or mammals;
_____Tissue Culture-for any projects involving growing eukaryotic tissues or cell cultures;
X Letter from institution where research was done or IJAS SRC, if an exception to the IJAS rules has been granted.

SIGNED
SIGNED

Student Exhibitor(s)
Sponsor *
*As a sponsor, I assume all responsibilities related to this project.

This Sheet Must Be Typed and This form must be displayed on the front of the exhibitors display board. It may be
reduced to half a sheet of paper 8.5 inches (vertical) X 5.5 inches (horizontal).

Microorganism Endorsement
The Illinois Junior Academy of Science

These rules will be strictly enforced for the State Science Exposition.
No region should send a project to the State Exposition that does not meet these regulations.
Students and sponsors doing a microorganism project must complete this form. The signature of the student or students and
the sponsor indicates that the project was done within these rules and regulations. Failure to comply with these rules will
mean the disqualification of the project at the state level. This form must follow the Safety Sheet in the project paper and on
the project board.
1. This area of science may involve many dangers and hazards while experimenting. It is the sole responsibility of all teacher(s)/sponsor(s) to teach students proper
safety methods and sterile techniques.
2. The Illinois Junior Academy of Science prohibits the use of primary or secondary cultures taken from humans or other vertebrate animals in any project because
of the danger from unknown viruses or other disease-causing agents that may be present. Pure cultures of microorganisms known to inhabit vertebrate animals may
be obtained from reputable suppliers and used in proper settings.
3. Microorganism experiments must be conducted in a laboratory such as science classroom or research facility.
4. Projects involving viruses and recombinant DNA should be done with the help of a professional and should comply with the National Institutes of Health (NIH)
Guidelines unless the project is limited to a kit obtained from a legitimate supply house.
5. All cultures should be destroyed by methods such as autoclaving or with a suitable NaOCl (bleach) solution before disposal.

Complete all boxes of the following chart.

DH5 Escherichia coli

Name of the reputable

The University of Chicago Biological Sciences Collegiate Division, originated from a reputable
organization *Not derived from a Primary or Secondary Culture

Method of disposal of
the organism(s) being used.

Disposal method included autoclaving, bleaching, and sanitizing following sterile techniques

List the location where the

The University of Chicago in the Biological Science Collegiate Division

Describe the use of

Microorganisms were treated with rifampicin and made antibiotically resistant

Other precautions taken

Secure place where the bacteria was taking, a safety course must be passed
A sanitation course also had to be passed.

The signatures of the student or students and sponsor below indicate that the project conforms to the above rules of the
Illinois Junior Academy of Science.

(Sponsor)*

(Student)

(Date)

(Student)

*As a sponsor, I assume all responsibilities related to this project.

This Sheet Must Be Typed

This form must be displayed on the front of the exhibitors display board. It may be reduced to half a sheet of paper
8.5 inches (vertical) X 5.5 inches (horizontal).
Check box if exception/approval letter from an institution where research was done, or the IJAS SRC is required and attached.