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LS14Mutations - Lecture 14 Genetic mutation Multiple levels...

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1 Lecture 14: Genetic mutation Multiple levels of gene regulations Different types of mutations Effects of mutations to gene expression and functions What cause cancer? Regulation of gene expression at multiple levels
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2 1. Nuclear pore and nuclear transport Nuclear pore structure contains 50-100 proteins called nucleoporins. In the cell, molecules smaller than ~60kD can diffuse freely through the nuclear pores, but those larger than 60kD cannot diffuse and they need selective transportation in or out of the nucleus. NLS: N uclear L ocalization S ignal of nuclear proteins Large nuclear proteins contain NLS. There are many different NLS sequences, often rich in basic amino acids. The SV40 NLS sequence, PKKKRLV , was the first one characterized. The NLS of a nuclear protein (X) can be fused, using recombinant DNA method, to a cytoplasmic protein (Y) to force the recombinant Y move into the nucleus Native pyruvate kinase is a cytoplasmic protein The pyruvate kinase fused to SV40-NLS is now in the nucleus
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