Attachment VI - Page 6 
2. General biological characteri sties, including genetics 
3. Pathological, ecological, and physiological traits 
4. Document the means and sensitivity of the techniques by which 
the presence or the absence of donor organism will 
be assessed in the final population of microoorgani sms to 
be released into the environment 
B. Genetic manipulations: describe the method(s) by which the 
DNA vector has been constructed and introduced. Include diagrams. 
1. Describe the nature of the DNA inserted or modified 
by the genetic manipulations. Document the amount of any donor 
DNA in excess of the amount required for the minimal effective 
genetic alteration in the final construction. Predict whether 
any changes in pathologic, ecologic, physiologic or genetic 
properties of host organism would be anticipated as the result 
of the donor DNA. 
2. Describe the amount of any vector DNA remaining in the final 
construction. Is any effect anticipated from this DNA? 
3. Describe the method of introduction of the DNA and selection 
altered organisms. 
a. If mi croorgani sms are used to introduce vectors or are 
vectors themselves, 
i. Indicate how they compare with wildtype strains. 
ii. If disarmed pathogens are used to transmit the 
vector, indicate factors that will most 
likely prevent these microorganisms from 
regaining or acquiring pathogenic potential. 
iii. If the vector is likely to survive independently of 
the host(s), refer to this possibility and provide any 
available data to assess the probability of transfer 
likely to other organisms 
iv. If a microorganism will only be used to introduce 
a vector (s) or are vectors themselves, document the 
means and sensitivity of the techniques by which the 
presence or the absence of the microorgani sm will be 
assessed in the final construction to be released 
D. Properties of the modified organism 
1. Describe anticipated properties of the final population of 
engineered organisms, with emphasis on the desired effect 
and any other possible pathologic, ecologic, physiologic or 
genetic effects. 
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