BIOINFORMATICS EXAM 2
protein sequence and
structure prediction-
overview test questions and
answers
why proteomics?
microarrays are easier and more established,
proteins are functional agents of the genome
ELISA (enzyme-linked immunosorbent assay)
a blood test used to screen for the presence of HIV
antibodies
elisa steps 1.
have a control and disease protein A mix
elisa steps 2.
plate of control and experiment
elisa steps 3.
add antibody
1. antibody and fluorophore
2. add primary antibody against protein A
to get antigen of protein A
introduce into the animals, collect antibodies from
the spleen
protein bioinformatics
, helps characterize protein sequences in silico and
allows prediction of protein structure and function
patterns of conservation in sequences allow us to
determine
which residues are under selective constraints
protein sequence alignment
pairwise alignment and multiple sequence
alignment
UNIPROT
is the most powerful database
3 steps to most proteomics experiments
-preparation of a complex protein mixture
-separation of protein mixture
-characterization of proteins within mixture
1.sample collection
controlled conditions, prevention of contamination
and degradation, difficult to purify proteins
separation techniques
-2D gel electropheresis- isoelectric focusing
-2d gel electropheresis- polyacrylamide gel
-liquid chromatography
2D-gel electrophoresis-isoelectric focusing
separation of proteins on basis of isoelectric point,
proteins migrate through pH gradient until their
overall charge is neutral
what is the isoelectric point?
protein sequence and
structure prediction-
overview test questions and
answers
why proteomics?
microarrays are easier and more established,
proteins are functional agents of the genome
ELISA (enzyme-linked immunosorbent assay)
a blood test used to screen for the presence of HIV
antibodies
elisa steps 1.
have a control and disease protein A mix
elisa steps 2.
plate of control and experiment
elisa steps 3.
add antibody
1. antibody and fluorophore
2. add primary antibody against protein A
to get antigen of protein A
introduce into the animals, collect antibodies from
the spleen
protein bioinformatics
, helps characterize protein sequences in silico and
allows prediction of protein structure and function
patterns of conservation in sequences allow us to
determine
which residues are under selective constraints
protein sequence alignment
pairwise alignment and multiple sequence
alignment
UNIPROT
is the most powerful database
3 steps to most proteomics experiments
-preparation of a complex protein mixture
-separation of protein mixture
-characterization of proteins within mixture
1.sample collection
controlled conditions, prevention of contamination
and degradation, difficult to purify proteins
separation techniques
-2D gel electropheresis- isoelectric focusing
-2d gel electropheresis- polyacrylamide gel
-liquid chromatography
2D-gel electrophoresis-isoelectric focusing
separation of proteins on basis of isoelectric point,
proteins migrate through pH gradient until their
overall charge is neutral
what is the isoelectric point?
