|Project Title||Investigation of Bacillus thuringiensis- endotoxin Binding to Midgut Receptors of Asiatic Corn Borer Ostrinia furnacalis|
|Project Type||Contained Use (Laboratory)|
|Name of Institution||National Institute of Molecular Biology and Biotechnology (BIOTECH),University of the Philippines Los Banos, College, Laguna,049-536-2721; 536-2724|
|Project Leader(s)||Edwin Alcantara, University Researcher I|
|Experimental Facility/Site||BIOTECH, Philippines|
|Purpose / Objectives||1. To identify Cry toxins with high binding affinity to midgut receptors of O. furnacalis.
2. To quantify receptor binding and determine which component of the binding process is critical to a successful Cry toxin- receptor interaction.
3. To develop a molecular approach to improve receptor binding of Cry toxin to O. furnacalis midgut receptors.
4. To determine the role of midgut receptors in the resistance of O. furnacalis to Bt toxin.
|Biosafety measures||1. Imported and locally transformed E. coli harboring cloned cry genes will be stored in glycerol at –80ºC freezer.
2. All aseptic work will be done in a laminar flow hood.
3. Growing of recombinant E. coli cells will be done in enclosed shakers with properly regulated speed and temperature.
4. All glasswares and solid and liquid wastes coming from the production of cloned B. thuringiensis endotoxin will be sterilized at 121ºC for 20 minutes in an autoclave.
5. Only researchers with experience in microbiological aspect techniques will be hired in the project.
6. Insect rearing cages with adequate containment will be provided for the rearing of B. thuringiensis toxin resistant insects. A separate rearing room with padlock will be provided for this purpose.
7. Discarded insects will be destroyed by autoclaving in 121ºC for 20 minutes.
|Conditions for Approval||In the induction of cry toxin resistance in O. furnacalis, cognizant of the importance of conserving the pest population’s susceptible gene pool the NCBP therefore enjoins the UPLB- IBC to closely monitor this resistance selection activity to ensure that the O. furnacalis strains selected for resistance are not allowed to reproduce outside the containment facilities|
|Date of Approval||1998-09-15|
|Regulated Materials||cry1Aa, cry1Ab, cry1Ac, cry1B, cry!C, cry1F, cry11A, and cry9 genes cloned into pKK223- 3 and transformed into E. coli J.M. 105|
|Source||Bacillus Genetic Stock Center, Ohio State University|
|Subject||Request of Dr. Agnes Zamora for permit to import cry1Aa, cry1Ab, cry1Ac, cry1B, cry!C, cry1F, cry11A, and cry9 genes cloned into pKK223- 3 and transformed into E. coli J.M. 105|
|Requesting Party||Dr. Agnes Zamora, Chairman, UPLB- IBC|
|Action of NCBP||Approved for Implementation
For issuance of permit to import cry1Aa, cry1Ab, cry1Ac, cry1B, cry1C, cry1F, cry11A, and cry9 genes cloned into pKK223- 3 and transformed into E. coli J.M. 105 explants of transformed bananas by the Plant Quarantine Services, Bureau of Plant Industry.