In my experimental design, I aim to track Trichosporon sp. fungus in lung tissue. To achieve this, I plan to utilize the actin gene promoter of Cryptococcus neoformans (CnPactin) and the termination sequence of the Aspergillus nidulans trpC gene. I will construct a cassette containing enhanced green fluorescent protein (eGFP) by incorporating the CnPactin promoter upstream of eGFP and the trpC terminator downstream of eGFP. This cassette will then be applied to the NAT-1 gene. For transformation, I intend to use EHA105 Agrobacterium. Any insights or suggestions regarding this approach and how to have the sequences of CnPactin, trpC, and NAT-1 would be greatly appreciated. Thank you.
Amina Abid
- Osama Sweef
- Many sequences are available in public databases like GenBank or specialized repositories for promoter and terminator sequences.
- Sequence Retrieval Services, Sequence Synthesis.
- Once you have obtained the sequences for the CnPactin promoter, trpC terminator, and NAT-1 gene, you can use molecular biology techniques such as PCR amplification, gene synthesis, or plasmid extraction to clone them into your expression cassette containing the eGFP gene. Make sure to verify the sequences through sequencing before proceeding with your experiments.
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