Q.3. Answer-
1. The change in one amino acid to another amino acid with same properties in protein is conservative missence mutation arises by the point mutation.
2. Here the tumor necrosis factor binding with adalimumab shows that glutamate in the light chain variable region enhances its binding activity. So that we want to change aspartate to glutamate in light variable chain .
3. The codon for aspartate is GAC and codon for glutamate is GAG. So that by creating point mutation at third nucleotide in aspartate codon we can code for glutamate. In GAC codon "C" is substituted with "G" and codon becomes GAG. This change from "C" to "G " is transversion mutation where pyrimidine is changed to purine.
4. This transversion mutation can be done by treating with chemical mutagen like alkylating agents or ionizing radiation.
VOLLES U LULUCULUULS. 3) Suppose that structural analysis of adalimumab binding to its target tumor necrosis!...
please give a detailed step by step laboratory
procedure. thank you.
3) Suppose that structural analysis of adalimumab binding to its target tumor necrosis factor suggests that hanging one amino acid in the light chain variable region from Asp to Glu will enhance binding activity. Describe in conceptual detail how you will create a mutation that changes the codon GAC to GAG in the light chain coding sequence.