19 Name of line: Nkx2-5:GFP
a. General Information:
1. Genetic background? Wild type X. laevis
2. Breeding history? F1 generation heterozygote (hemizygote) for transgene.
3. What is the inserted DNA? 8 kb of promoter of the X. laevis Nkx2-5 gene driving expression of GFP (variant mt5). Mohun Lab Plasmid #2147. Contact Tim Mohun for the sequence as it differs from the published NCBI AF283102. The promoter of the second laevis Nkx2-5 allele was used here. It proved stronger than the originally published one (Sparrow et al).
4. Are there any specific PCR reactions or genomic Southern blots that can be used to identify the line unequivocally? (please supply sequences or probe details). See sequence.
5. Is the insertion site and/or copy number of the transgene known? No.
6. Expression pattern of transgene? Early heart field, mesoderm and pharyngeal endoderm. Spleen.
7. Do the animals require special treatment during routine maintenance or breeding? Select the best fluorescing animals at developmental stage 25. Continue to monitor to stage 45.
8. Can these animals be made freely available to the research community? (please detail any restrictions necessary) MRC-NIMR Depositor MTA required.
b. Risk assessment of transgenic Xenopus (required under UK law):
1. Please provide a short description (i.e. function in donor organism) of the inserted genetic material. (example: labels interneurons) Nkx2-5 gene promoter drives expression of GFP in the early heart field.
2. Please describe the method used to insert the foreign DNA. Sperm nuclear injection transgenesis.
3. What are the characteristics of the recombinant organism? Embryonic heart field fluoresces.
4. Does the recombinant organism potentially have harmful allergenic or toxic effects on humans? No.
5. Could the recombinant organism act as a human disease vector or reservoir? No.
6. Could the recombinant organism have an adverse effect on human health arising from change in behaviour or in physical nature? No.