Lab #8-9: Identification of Mutations

ATCG
ACCG
Stop codon: TAA, TAG, TGA
Single Nucleotide Polymorphism (SNP) (Point mutation)
Step 1: get your fasta sequence from NCBI database.
Get insulin nucleotide sequence
Step 2: Open BLAST tool and click on BLASTx program.
Step3: Perform BLASTx from nucleotide to protein
Step 4: See pairwise alignment and analyse the mutations
occurrence
Write the mutation in a format: for example,
H6R (Naming of Point Mutation)
3 things are needed:
For verification we have various tools:
1. Polyphen 2:
http://genetics.bwh.harvard.edu/pph2/
Expression decrease or increase
Probably damaged or Benign
2. Provean
http://provean.jcvi.org/seq_submit.php

3. MUpro
http://mupro.proteomics.ics.uci.edu/
 4. I-Mutant
http://gpcr2.biocomp.unibo.it/cgi/predictors/I-Mutant3.0/I-
Mutant3.0.cgi

5. PHD-SNP
https://snps.biofold.org/phd-snp/phd-snp.html
6. SIFT
https://sift.bii.a-star.edu.sg/www/SIFT_seq_submit2.html

>M57671.1 Octodon degus insulin mRNA, complete cds

GCATTCTGAGGCATTCTCTAACAGGTTCTCGACCCTCCGCCATGGCCCCGTGGATGCATCTCCTCACCGT
GCTGGCCCTGCTGGCCCTCTGGGGACCCAACTCTGTTCAGGCCTATTCCAGCCAGCACCTGTGCGGCTCC
AACCTAGTGGAGGCACTGTACATGACATGTGGACGGAGTGGCTTCTATAGACCCCACGACCGCCGAGAGC
TGGAGGACCTCCAGGTGGAGCAGGCAGAACTGGGTCTGGAGGCAGGCGGCCTGCAGCCTTCGGCCCTGGA
GATGATTCTGCAGAAGCGCGGCATTGTGGATCAGTGCTGTAATAACATTTGCACATTTAACCAGCTGCAG
AACTACTGCAATGTCCCTTAGACACCTGCCTTGGGCCTGGCCTGCTGCTCTGCCCTGGCAACCAATAAAC
CCCTTGAATGAG

>AAA40590.1 insulin [Octodon degus]

MAPWMHLLTVLALLALWGPNSVQAYSSQHLCGSNLVEALYMTCGRSGFYRPHDRRELEDLQVEQAELGLE
AGGLQPSALEMILQKRGIVDQCCNNICTFNQLQNYCNVP