Project Information

Reference Number 20120269
Project Title Cloning and Expression of Raw Starch Digesting-Amylase Genes from Saccharomycopsis Fibuligeria and Saccharomycopsis Bubodii for Direct Ethanol Fermentation
Project Type Contained Use (Laboratory)
Status Ongoing
Name of Institution University of the Philippines - Mindanao
Cooperating Institution UP Mindanao Research
Supervising IBC University of the Philippines-Mindanao
Project Leader(s)

Annabelle U. Novero, Ph.D.

Experimental Facility/Site UP Mindanao, Mintal Davao City
Purpose / Objectives

The primary aim of this project is to characterize the putative RSDA genes from Saccharomycopsisfibuligera and Saccharomycopsisbubodii 2066 and express them in E.coli. The project has the following specific aims:
a. To design primers based on the conserved regions (encompassing the entire gene) of the RSDA sequences deposited in the database;

b. To use primers designed in objective a, for the construction of a cDNA library;

c. To determine the sequence of the RSDA genes cloned in E. coli;

d. To register the DNA sequences in the databases; and

e. To determine protein expression of the RSDAs cloned in E. coli thus, proving their functionality.

Biosafety measures

The experimentation will be conducted in the Molecular Cloning Laboratory of UP Mindanao, a BL2 laboratory. Researchers who will conduct the study are given proper orientation and demonstration and are recommended by the laboratory director. They are also given awareness of the biosafety procedures. Access to the laboratory is limited and restricted at the discretion of the laboratory director especially when the experiment is in progress. Wearing of prescribe laboratory gown, safety goggles and gloves are observed. Wearing contact lenses, sandals or open-toed shoes are prohibited. To keep away from any chemicals, burners and candles or other laboratory equipment, long hair are tied back and jewelleries are removed. Working area is cleaned and disinfected before and after the experiment and spills are properly cleaned. Cabinets, refrigerators and ultralow freezers are provided for the storage of laboratory consumables and reagents. The laboratory is designed to have a sink for hand-washing and shower for emergency purposes only. Designation of risk areas are properly labeled and strictly imposed to the laboratory users. Biosafetiness of the equipment, glassware and chemicals are also provided. Material safety data sheets (MSDS) for each chemical reagent to be used will closely studied before use. Proper sterilization of laboratory wares and consumables used in the experiment are done using the autoclave machine. Autoclave is cleaned once a month. All contaminated liquid or solid wastes are decontaminated before disposal. All types of chemical reagents waste used especially products of plasmid cloning will be discarded in accordance with the Proper Guidelines on Health and Biosafety.

Conditions for Approval

All activities shall be conducted in the Molecular Cloning Laboratory of UP Mindanao, Brgy. Mintal, Tugbok District, Davao City. The proponent shall adhere as closely as possible to the schedule of activities reflected in the submitted Gantt chart. Any modification in the schedule of activities should be made with the concurrence of the UP Mindanao-IBC and the DOST-BC. A biosafety contingency plan must be submitted before the conduct of the experiment. The proponent shall inform which activities would require the presence of the UP Mindanao-IBC, DOST-BC and PEQS-BPI monitors. The proponent shall ensure that only the DOST-BC authorized personnel shall work on the project within the premises of the designated experimental facilities. The proponent shall ensure that stray animals are exluded from the experimental facilities while experiments are being conducted. In case of undue destruction of the experimental materials resulting from unauthorized entry of personnel or breach of containment of the laboratory facilities, the proponent must implement measures to prevent the inadvertent escape of any viable material within the facilities. The proponent and the supervising IBC shall be held accountable for the undue destruction of the experimental materials and the consequences that its inadvertent escape may cause to the surrounding environment. All viable materials within the experiment area shall be accounted for. The transport or movement of transgenic materials over a significant distance (i.e. from one laboratory/building/institution to another) shall not be allowed unless with the permission and under the supervision of the DOST-BC/PEQS-BPI and the UP Mindanao-IBC. Transgenic plant materials to be moved or transported must be placed in containers that are leak-proof and not easily broken (i.e. plastic bags), tightly sealed and properly labeled. These should then be placed in a secondary container to avoid accidental leakage or spillage (as per Appendix 2: Guidelines for Transport and Movement of Genetically Modified Organisms and Potentially Harmful Exotic Species of the “Guidelines on Planned Release of Genetically Manipulated Organisms (GMOs) and Potentially Harmful Exotic Species”, (NCBP Series No. 3, p. 42-44). Any additional requirement that the DOST-BC may impose, as necessary, during the experiment period should be complied with. The proponent shall submit through the IBC a terminal report 90 days after completion of the experiment. The proponent shall strictly observe proper disposal procedures of the viable materials.

Date of Approval (DD-MM-YYYY) 30-05-2012
Date of Completion (DD-MM-YYYY) N/A
Executive Summary N/A