Andrew D. Montecillo
Ma. Theresa Jonna A. Atienza
To determine the gene responsible for the AuNP production of PGPB through the aid of transposon mutagenesis
1. Proper laboratory attire (lab gown, closed shoes, gloves, etc.) will be worn at all times.
2. The correct containers will be used in conducting transposon mutagenesis experiments.
3. The isolates and other materials will be handled carefully to avoid spills.
4. Work surface will always be sterilized before and after every activity.
5. The researcher will wash and disinfect her hands after working with the isolates and before leaving the laboratory.
6. Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human consumption are not permitted in the laboratory.
7. All materials that were in contact with the isolates will be disinfected and/or decontaminated.
8. The cultures and containers will be decontaminated once the project has been terminated.
a) Project activities shall be conducted at the BL2 lsolatron Room at Molecular Genetics Laboratory of the University of the Philippines Los Banos - National lnstitute of Molecular Biology and Biotechnology (UPLB-BTOTECH).
b) The proponent shall adhere as closely as possible to the schedule of activities reflected in the submitted Gantt chart.
c) Any modifications in the schedule of activities shall be made with the concurrence of the UPLB-IBC and the DOST-BC.
d) Signage should be in place indicating that the facility is restricted/limited access.
e) The proponent must provide a logbook to record access of personnel to the experimental facility.
f) The proponent must take measures to ensure that only the DOST-BC authorized personnel are allowed inside the experimental facilities.
g) The experimental facilities must be kept under lock when not in use.
h) ln case of undue destruction of experimental materials resulting from unauthorized entry of personnel or breach of containment of experimental facilities, the proponent shall implement measures to prevent the inadvertent escape of any viable material within the facility.
i) The proponent and the supervising IBC shall be held accountable for the undue destruction of the experimental materials and the consequences that their inadvertent escape may cause to the surrounding environment.
j) The DOST-BC should be informed in advance of any visitations by unauthorized persons.
k) The DOST-BC shall be informed immediately of any intrusion by unauthorized persons.
l) The proponent shall strictly observe proper disposal procedures for all materials.
m) Movement of all materials will be done in compliance with all relevant biosafety and phytosanitary requirements of the Philippines.
n) Any additional requirement that the DOST-BC may impose, as necessary, for the duration of the experiment shall be complied with.
o) The proponent shall submit through the IBC a completion report within 90 days after completion of the experiment.
p) The IBC shall submit to the DOST-BC a report on the completion of this project, in the prescribed format.
One of the most exciting fields in nanotechnology is the synthesis of nanoparticles. These nano particles have broad range of uses from medicine to cosmetics. The synthesis of gold nanoparticles (AuNPs) gained attention due to its use in the field of medicine. Biological synthesis of AuNPs using microorganisms allows the production of homogeneous and monodispersed AuNPs. Some plant growth promoting bacterial (PGPB) isolates were found to synthesize gold nanoparticles in the 10-100 nm size range. However, the genese and pathway for AuNPs synthesis is not yet elucidated.
This project will utilize transposon mutagenesis to determine the gene/s responsible for the gold nano particles (AuNPs) biosynthesis of PGPB. Other advanced molecular biology techniques will also be employed in the study. The study involves the insertion of the plasmid pRL27 containing Tn5 from Escherichia coli to a PGPB isolate through becterial conjugation. This will be followed by the characterization of the transconjugants and the wild type via AuNP production assay. Genomic DNA extraction and polymerase chain reaction of kanamycin resistance gene, and the transformation of E. coli and selected bacterial strains by electroporation will also be performed. All the experiments will be performed at the Nano-Biotech Information and Translational Laboratory (NBITRL) in BIOTECH-UPLB from November, 2016 to March, 2017.
Public Information Sheet
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