APHIS Notification Procedures
Information needed to receive transgenic seeds from the Maize COOP:
Note: All of the construct information about RescueMu is included in the sample notification letter (below), so you will only need to cut and paste that information. For other transgenic stocks in our collection (e.g., from the Chromatin Functional Genomics Consortium, tdsg, or MAIZE CELLGENOMICS stocks), please request the construct information for each stock.
For requests from outside the USA:
The requester must inquire about any regulatory
requirements from their government, and let us know if we need to do
anything special.
For requests from within the USA:
In order to receive the shipment, you are required
to send a movement notification letter (see below for a sample) to
APHIS.
From outside Illinois, this is called 'interstate movement'; from
within
Illinois this is called 'intrastate movement'.
In order to grow these stocks under field, greenhouse or laboratory conditions, you will also need to submit a release notification. Movement and release notifications can be sent simultaneously, in the same letter. After review by APHIS and your state's officials, the notification will be acknowledged. See: Permits Notifications and Petitions for further information.
Note: a notification for movement should be acknowledged within 10 days of receipt by APHIS; and for release, within 30 days of receipt.
If you would like to keep your name and address confidential in the public release version of the notification, please ask APHIS personnel how to have this 'Personally Identifying Information (PII)' withheld from the public record.
Notifications can now be sent through APHIS's ePermits system. If you use the ePermits system for RescueMu stocks, you can copy/paste the information (in the sample) below for Section 6 (Description of Regulated Article). For other transgenic stocks, please ask us for a sample Notification to use as a template.
Alternatively, a notification letter may be submitted by mail and/or by FAX. However, since the signature on a notification letter submitted by FAX is not legally binding, it is required that confirmative copies with legally valid (original) signatures be submitted by mail (preferably by an overnight delivery system that guarantees and tracks deliveries). A template for writing a notification letter for RescueMu stocks is provided below.
Suggestion: It seems to be better if PIs rather than students/staff send the notification to APHIS.
There are also requirements for how you should keep
these seeds-- see http://www.aphis.usda.gov/biotechnology/
and http://www.access.gpo.gov/nara/cfr/waisidx_05/7cfr340_05.html
[Container requirements for regulated articles]. Regulations also specify
that you must prevent any mixing of transgenic stocks with non-transgenic
ones and prevent any accidental escape of seeds or plants from your storage facility.
APHIS notification example for receiving RescueMu stocks from the Maize COOP:Sample Notification: (see: Permits Notifications and Petitions)Please modify this sample to suit your individual needs.
Permits Branch
Biotechnology Permits
Biotechnology, Biologics and Environmental Protection
4700 River Road
Riverdale, MD 20737-1228
Re: Notice of Environmental Release and Interstate Transfer
1. Name, Address, Telephone and Email of Applicant:
2. Introduction Type:
3. Applicant Reference Number: [assign number for your internal purposes and to track progress of the acknowledgement]
4. Confidential Business Information Verification (CBI):
5. Regulated Article (Everything in green text is the information you need to supply):
Scientific Name:
Zea mays
Common Name:
maize
Cultivar and/or Breeding Line:
(optional)
6. Phenotypic Designation
1. Phenotypic Designation Name: (optional)
Identifying Line(s): (optional)
Construct(s):
pMuRep2-pAHC20
Mode of Transformation:
Biolistic
Phenotype(s)
MG - Colored sectors in seeds
Genotype(s)
Gene(s) of Interest
Promoter: 35S 5'
from
Cauliflower mosaic caulimovirus
-
initiation of transcription
Transcription Factor: Lc (R) (Leaf color anthocyanin)
from
Zea mays - - Lc (R) is a transcription factor of the R gene family in maize which is required for anthocyanin production. Serves as a marker of transposon excision where transposition
of RescueMU from the Lc::RescueMu transgene leads to restoration of pigmentation.
Transposon: RescueMu2
from
Zea mays - - RescueMu is a modified Mutator1 transposon (Mu1) containing bacterial pBluescript plasmid and transformed into maize to permit mutagenesis and subsequent recovery of mutant
alleles by plasmid rescue.
Marker Gene: 200 bp marker DNA
from
Sinorhizobium meliloti - To distinguish between different RescueMu mobile elements, a unique 200 bp tag from the S. melioti nodulin gene has been inserted into the RescueMu2 transposon
to permit easier mutant allele-transposon cosegreation analysis. No gene is expressed in transgenic maize as a result of this marker.
Terminator: rbcS 3' end from rbcS-E9 gene
from
Pisum sativum - Terminator sequence of pea rbcS-E9 gene which in this case halts transcription of the maize Lc (R) anthocyanin transcription factor.
Vector Sequence: pBluescript
from
Escherichia coli - pBluescript (commercial cloning vector plasmid from Stratagene) contains a bacterial origin of replication and an antibiotic resistance gene (ampicillin) to permit
plasmid rescue.
Selectable Marker
Promoter: Ubiquitin
from
Zea mays - Initiation of transcription
Enhancer: Ubiquitin 1 intron
from
Zea mays - Enhancer of transcription
Gene: phosphinothricin acetyltransferase (bar)
from
Streptomyces hygroscopicus – Bialaphos resistance (bar) gene from pAHC20 plasmid encoding the phosphinothricin acetyl transferase (pat) enzyme which confers glufosinate tolerance, which
in this case is used as a selectable marker to indicate the stable insertion of RescueMu.
Terminator: nos 3'
from
Agrobacterium tumefaciens - Termination of transcription
2) Phenotypic Designation Name:
Identifying Line(s):
Construct(s):
pMuRep3-pAHC20
Mode of Transformation:
Biolistic
Phenotype(s)
MG - Colored sectors in seeds
Genotype(s)
Gene(s) of Interest
Promoter: 35S 5'
from
Cauliflower mosaic caulimovirus - initiation of transcription
Transcription Factor: Lc (R) (Leaf color anthocyanin)
from
Zea mays - - Lc (R) is a transcription factor of the R gene family in maize which is required for anthocyanin production. Serves as a marker of transposon excision where transposition
of RescueMU from the Lc::RescueMu transgene leads to restoration of pigmentation.
Transposon: RescueMu2
from
Zea mays - - RescueMu is a modified Mutator1 transposon (Mu1) containing bacterial pBluescript plasmid and transformed into maize to permit mutagenesis and subsequent recovery of mutant
alleles by plasmid rescue.
Marker Gene: 400 bp marker DNA
from
Sinorhizobium meliloti - To distinguish between different RescueMu mobile elements, a unique 400 bp tag from the S. melioti nodulin gene has been inserted into the RescueMu2 transposon
to permit easier mutant allele-transposon cosegreation analysis. No gene is expressed in transgenic maize as a result of this marker.
Terminator: rbcS 3' end from rbcS-E9 gene
from
Pisum sativum -
Terminator sequence of pea rbcS-E9 gene which in this case halts transcription of the maize Lc (R) anthocyanin transcription factor.
Vector Sequence: pBluescript
from
Escherichia coli - pBluescript (commercial cloning vector plasmid from Stratagene) contains a bacterial origin of replication and an antibiotic resistance gene (ampicillin) to permit
plasmid rescue.
Selectable Marker
Promoter: Ubiquitin
from
Zea mays - Initiation of transcription
Enhancer: Ubiquitin 1 intron
from
Zea mays - Enhancer of transcription
Gene: phosphinothricin acetyltransferase (bar)
from
Streptomyces hygroscopicus – Bialaphos resistance (bar) gene from pAHC20 plasmid encoding the phosphinothricin acetyl transferase (pat) enzyme which confers glufosinate tolerance, which
in this case is used as a selectable marker to indicate the stable insertion of RescueMu.
Terminator: nos 3'
from
Agrobacterium tumefaciens - Termination of transcription
7. Introduction:
Release:
NUMBER OF STATES AND SITES: [your state] (1)
*[your county]
[amount, e.g., number of acres]
Interstate Transfer:
ORIGIN: IL DESTINATION: [your state (Your County)]
1) FROM: Champaign County, IL, as <1 lb of corn seed
TO: [Your organization, your county, your state]
8. Certification: I certify that the regulated article will be introduced in accordance with the eligibility criteria and the performance standards set forth in 7 CFR 340.3. The above information is true to the best of our knowledge. If there are any changes, we will contact APHIS.
Signature ____________________________ Date: [Date]
[ Name ], [ title]
[organization]