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Among the most widely applied and most powerful of these techniques is forward genetic screening, in which (usually random) alterations are made in the genome of an organism, and the resulting phenotypes are studied to divine the function of the altered genes.
To test the function of the altered Env proteins, mutant envB genes were inserted upstream of the internal ribosome entry site (ires)- enhanced green fluorescent protein (eGFP) cassette in the murine leukemia virus-based retroviral vector pCMMP.IRES.eGFP [32].
The function of the altered genomes was then validated by genome transplantation and phenotypic characterization of the transplanted cells.
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Amino acid substitutions may also affect the function of the protein, altering optimal ranges for temperature, pH, or substrate concentration.
For example Kimchi-Sarfaty and colleagues found that a single-nucleotide polymorphism (SNP) which does not alter the coding sequence has a drastic effect on the function of the protein without altering the mRNA or protein levels [ 17].
Both CCND1 and COMT polymorphisms have been previously shown to have an impact on the function of the protein products, altering their overall enzymatic activity in the cell.
Conventional theory regarding the effect of polymorphisms on CYP1A1 suggests that the variants affect the function of the enzyme by altering the level of gene expression or the mRNA stability, although the results in the literature appear to be contradictory [ 98].
Lysine residues contribute to the net positive charge of ANT1 that is critical for transport of ADP/ATP, and acetylation of these residues could affect the function of the protein by altering charge-based interactions.
The normal cellular function of the altered NDRG1 may not be maintained.
Here, we assumed that the function of the gene was not altered by its elongation or contraction.
25, 27 In the context of elevated blood glucose, the function of the vascular endothelium is altered in ways that promote atherosclerosis (reviewed in Bornfeldt and Tabas 62).
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