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Do you HYPHY with (Data)Monkey !!

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HyPhy, acronym for Hypothesis Testing Using Phylogenies ( was written & designed by Kosakovsky Pond and workers to provide likelihood-based analyses on molecular evolutionary data sets and help detect differential rates of variability within a coding sequence datasets. It is freely available, has a Graphical User Interface and can be used by anyone with or without much computer language or programming exposure.

It was earlier presumed that substitution rates were uniform over an alignment of homologous DNA/Protein sequences but many workers studying molecular evolutionary processes influencing rates and patterns of evolution negated this presumption with quite a lot of data and this is especially true for highly evolving gene family datasets and for viral genomes. Natural selection takes place at different domains/regions/sites which are under positive, negative or neutral selection pressures. Positive selection originates with more of non-synonymous substitutions in a protein coding sequence influencing the fitness advantage (protein structure and function) of an organism whereas negative selection takes place with more of synonymous substitution in a protein coding sequence leaving the amino acid sequence or protein structure and function unchanged. A neutral evolution is said to be taking place when the non-synonymous substitutions do not affect the protein structure and function and rate of non-synonymous substitutions. The rate of synonymous and non-synonymous substitutions is given by dS and dN respectively. In the case of neutral evolution, dS and dN are observed to be in equilibrium. Accordingly, the ratio of dN/dS given by ω=β/α (also referred to as dN/dS) has become a standard measure of selective pressure. The total ω  for a sequence alignment is referred to as Global ω. Global ω with a value of approximately 1 signifies neutral evolution, below 1 suggests negative selection whereas ω more than 1 implies positive selection. To start with the analyses, all one needs is, a suitable codon substitution model as detected by MODELTEST program (available online), a nexus formatted sequence alignment file (must be codon data file) and a Maximum Likelihood tree of the data.

Datamonkey is a web interface ( which uses HyPhy batch files to execute most of its tools and packages for the computational analyses. This web interface can be used for estimating dS and dN over an alignment of coding sequences and also for identifying codons and lineages under selection. It also provides “state of the art” tests of codon based models to infer signatures of positive Darwinian selection by comparing rates of synonymous (dS) versus non-synonymous (dN) mutations even in the presence of recombination. It actually reports ω (=dN/dS) using a variety of evolutionary models. Apart from this, Datamonkey also offers a number of packages such as GARD, SLAC, REL, FEL, EVOBLAST etc. These will be discussed in the next issue. Keep reading!!


A comprehensive list of references on the article are available upon request to the author ([email protected])

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Dr. Pant is a researcher with keen interest in software driven analysis of DNA/Protein sequence data for taxonomic, phylogenetic and other homology based studies. Currently he is involved in understanding Microbial diversity using Next generation sequencing approaches and Analysis of sequence and metagenomic datasets using computational biology approaches. He is presently engaged with undergraduate teaching as an Assistant Professor in University of Delhi

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