Homology
Functions
Pairwise sequence alignment
SCORES
identity
mismatch
gap penalty
opening - inroduce gap
extension - elongate gap
scores
raw scores (calculate from substitution matrix)
bit scores (normalized scores)
SCORING MATRICES
PAM
PAM1
probability of occurance
+
percent accepted mutations
=
mutation probability matrix
at least 85% identical
one PAM (unit of evolution) =
1% amino acids changed between two proteins
Percent Accepted Mutations (Dayhoff)
global alignment (closely related protein)
X% divergence
BLAST
Blocks Substitution Matrix (Henikoff)
local alignment (distantly related protein)
X% identical
Smith-Waterman local alignment
four component for BLAST search
CHOOSE sequences
SELECT BLAST program
blastp
blastn
blastx
tblastn
tblastx
CHOOSE database
nr
Month
Swissprot
RefSeq
Pdb
pat
Subtopic
pnv_nr
CHOOSE optional parameters
different for every blast program
organism
turn filtering on/off
substitution matrix
expect value
Extreme Value Distribution (EVD)
E = Kmn e-λS
word size
output format
BLAT
BLAST-like alignment tool
500 times faster than BLAST
design to rapidly align longer nucleotide sequence
more than 95% similarity
MUTATIONS
can happens by
substitution
insertion
deletion
Mutability
high mutability
Asn, Ser, Asp, Glu
low mutability
Tyr, Phe, Leu, Cys, Trp