Enhanced Transfection of Target DNA.

Topics: Stem Cells

Essay, Pages 2 (268 words)

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All four of the wells were condensed with I Pl of an enhancer and added too lipid-based transaction reagent Effected to allow the target DNA to be introduced into the Hell cells. The cells containing . 45 ml Of transaction complexes and complete growth medium were incubated for two days and then replaced With I ml Of HOBS and incubated for another hour. Approximately 14. 13 Pl of glorification documentation diluted in PBS was added to one of the two wells containing GAP DNA (Figure 1 B) and a second well containing GAP-KGB DNA (Figure 1 D).

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Figure 1 Shows the GAB-KGB DNA lacking documentation and figure 1 A displays the Hell cells with just GAP DNA The Hell cells with GAP DNA and documentation (Figure 1 B) fluoresced throughout the entire cell including the nucleus most likely caused by the small size of the fluorescent protein used. However, when GAP-KGB DNA that lacked the documentation was analyzed, the entire cell fluoresced similarly except for the nucleus (Figure 1 C), indicating that the protein does not contain a nuclear localization signal.

The nucleus is fluoresced at a high intensity when documentation was added to the

GAP-KGB DNA containing cells (Figure I D) because the protein was capable of passing through to the nucleus. When the glorification hormone documentation binds to KGB, a conformational change causes neighboring proteins to dissociate and reveal the nuclear localization signal, therefore fluorescing the nucleus These results conclude that the GIBE-KGB DNA was successfully transferred into the Hell cells and through the addition of documentation, the glorification receptor protein with an activated glorification-binding domain localized to the nucleus following a conformational change.