As part of my Honours thesis in Biochemistry, I generated an expression construct containing the Dictyostelium homolog of LITAF and transformed it into Dictyostelium to conduct immunofluorescence localization studies of LITAF at different stages of growth and development. This will not only allow the tracking of changes in the localization of wildtype LITAF but also of its human mutant forms already developed by Dr. Brunetti’s laboratory. Thus, the identification of LITAF localization pattern in Dictyostelium throughout its development will offer valuable insight into the protein’s function. Furthermore, I intend to also develop a LITAF gene knockout in Dictyostelium using various molecular techniques such as RNA interference, PCR amplification, cloning, transformation and Southern blotting. A gene knockout study will further elucidate LITAF function by allowing the identification of abnormalities during growth and development due to its gene inactivation. Dr. Robert Huber’s laboratory has already successfully conducted a gene knockout in Dictyostelium of Cln3, a protein associated with Batten disease.8 Thereby, his lab offers valuable experience and expertise in this methodology essential to the success of this
As part of my Honours thesis in Biochemistry, I generated an expression construct containing the Dictyostelium homolog of LITAF and transformed it into Dictyostelium to conduct immunofluorescence localization studies of LITAF at different stages of growth and development. This will not only allow the tracking of changes in the localization of wildtype LITAF but also of its human mutant forms already developed by Dr. Brunetti’s laboratory. Thus, the identification of LITAF localization pattern in Dictyostelium throughout its development will offer valuable insight into the protein’s function. Furthermore, I intend to also develop a LITAF gene knockout in Dictyostelium using various molecular techniques such as RNA interference, PCR amplification, cloning, transformation and Southern blotting. A gene knockout study will further elucidate LITAF function by allowing the identification of abnormalities during growth and development due to its gene inactivation. Dr. Robert Huber’s laboratory has already successfully conducted a gene knockout in Dictyostelium of Cln3, a protein associated with Batten disease.8 Thereby, his lab offers valuable experience and expertise in this methodology essential to the success of this