IBC # 00-000

Recombinant DNA in Pathogens

(This form covers both Animal and Plant pathogens):

Section 1: General Information

Protocol Title:

Principal Investigator:

SECTION 2: Recipients of recombinant dna
This pathogen will be: / Special Characteristics / For the introduction of Viral Genomes Only
Recombinant Pathogen
(Recipient of recombinant DNA) / Will the modified pathogen be replication competent? (Yes or No)
If not competent, how will this be verified? / Used in cultured cells/tissues
(Yes or No) / Used with a helper virus or helper plasmid? (Yes or No)
If yes, specify / Introduced into non-human vertebrates, invertebrates or plants
(Yes or No) / Known antibiotic resistance, natural isolate, etc. / % of viral genome to be used:
<50%, <66%, >66%, 100%
(*Note: all members of the same family are considered identical)

*Note: An existing spreadsheet including the specific information requested above may be inserted or submitted with your application -as APPENDIX B- in place of the above table.

SECTION 3: Modified dna

Recombinant DNA categories to be transferred to pathogen(s) listed above (Check all that apply):

Protein-coding (marker/reporter) Protein-coding (other)

Non-coding Catalytic RNA miRNA Antisense RNA

Double-stranded RNA/siRNA Complex sample (genomic library, etc) Other

Method/Type of transfer into pathogen (Check all that apply)

Genomic/Chromosomal insertion Plasmid/Episomal Virus Transposon

Recombination Nanoparticles Biolistics Molecular cloning Other

Briefly describe how recombinant DNA molecules will be introduced into each pathogen genome:

*Note: An existing spreadsheet including the specific information requested below may be inserted or submitted with your registration -as APPENDIX C- to satisfy the information required in the following 3 tables.

For each protein-coding gene (except marker/reporter genes) or catalytic RNA, provide:

Name / Source of gene or catalytic RNA (e.g., name of native organism or “synthetic”) / Known or suspected function(s) / Pathogen DNA is being inserted into (if more than 1 pathogen is listed in Section 2)

For each non-coding segment, miRNA/dsRNA/siRNA or complex sample, provide the source organism (for synthesized molecule, provide the source of the native sequence):

Type / Source Organism / Pathogen DNA is being inserted into (if more than 1 pathogen is listed in Section 2)

If "Other", please explain the type and source of recombinant DNA:

Type / Source Organism / Pathogen DNA is being inserted into (if more than 1 pathogen is listed in Section 2)

Y N

Is this the deliberate transfer of recombinant DNA derived from a eukaryotic viral genome into a pathogen genome?

If yes, please list all sources of viral-derived recombinant DNA in the table below:

SECTION 4: Modified Viral DNA/ Viruse(s) used as DNA source(s)
Viral genome
(Source of recombinant DNA) / Is the viral-derived recombinant DNA to be transferred replication competent itself? / % of viral-derived recombinant DNA to be transferred into the target pathogen
(Note: all members of the same family are considered identical) / If <66% of a viral genome will be introduced, will a Helper be used to restore infectivity of the source virus following introduction into the target pathogen? / Is there a reasonable expectation that complementation or recombination could restore infectivity of the source virus once introduced into the target pathogen? / Pathogen DNA is being inserted into (if more than 1 pathogen is listed in Section 2)

*Note: The above table concerns sources of recombinant DNA (only viral in nature), whereas the opening table concerned recipients of recombinant DNA. In some cases such as viral chimeras, this distinction may be less clear, and in this case the majority sequence (in terms of nucleotides) should be referred to as the 'recipient', with the minority sequence as 'source'.

For each recombinant viral genome used as a source of recombinant DNA to be transferred into a pathogen and if less than 100% of the source viral genome is used- provide a brief summary as to the potential for the fraction of the viral genome being utilized to lead to a productive infection:

SECTION 5: Effect of modification(s) on the pathogen

Y N

Does any of the above named recombinant DNA represent the deliberate transfer of a drug (or other type of) resistance trait into a microorganism that is not known to occur naturally?

If yes, list antibiotic / resistance genes which will be inserted and any new traits conferred to any/all organisms:

Y N

Could such a transfer compromise the use of a drug (or other compound) to control disease agents in humans, veterinary medicine, or agriculture?

Provide a brief explanation justifying your response:

Y N

Is there a reasonable expectation that this modification will enhance the pathogenicity of the recipient or source pathogen?

If yes, explain and describe the expected phenotype (provide references if applicable):

Y N

Is there a reasonable expectation that this modification will extend the host range of the agent?

If yes, explain and describe the phenotype (provide references if applicable):

Y N

Is there an intentional release (of recombinant DNA-containing pathogens) into

the environment planned for this program?

If yes, briefly describe how these experiments will be conducted; and describe the anticipated effect on human or animal health as well as the environment:

*Note: The process and act of releasing genetically modified microbes into the environment may be regulated by USDA-APHIS and/or the FDA. If you have answered, "Yes" to this question, your application will not be considered complete unless the appropriate permit application and a copy of the approved permit accompany this application. See http://www.aphis.usda.gov/biotechnology/index.shtml for more information.

Recombinant DNA in Pathogens 1 of 4 Rev. 8/22/13