Summary: In this paper we introduce version 2.0 of CheckAlign, an open source application oriented to bioinformatic analyses based on information theory and the Shannon-Weaver algorithm.
Remarks: The update consists in the following: (a) a comprehensive source code optimization to adapt CheckAlign to the new versions and updates of the most common operating systems; (b) the implementation of an additional utility for estimating diversity index measures for one or more biological or molecular samples based on the proportional abundance of species in the samples.
Availability: CheckAlign 2.0 is an open source software project available both as an online server and as standalone software. The server is publicly accessible at [URL1]. Software and source code can be downloaded from [URL2].
Information theory is a branch of applied mathematics involving the quantification of information. Soon after its inception, information theory  was applied to distinct areas of biological research. Of significant interest was the sequence logo methodology [2,3], which offers graphical representations of DNA or protein multiple alignments, providing a statistically relevant visualization of the consensus of a set of sequences, their common information content, and the frequency of all possible DNA and amino acid states per alignment position. In 2008, taking this methodology into consideration, we designed the first version of CheckAlign, a logo-maker application that builds sequence logo representations online and on PCs. In this paper, we present an update of CheckAlign to version 2.0. The update is based on a source code optimization to adapt the logo maker to the updates of the most common operating systems and an implementation of the Shannon-Weaver algorithm  to characterize species diversity in a community. This means that CheckAlign now focuses not only on alignment analyses, but also on other ecological or molecular analyses concerning the rarity and commonness of species in a community. The ability to quantify diversity in this manner is useful for understanding the structure of a biological community. This is valid for evaluating the diversity of an ecological community of organisms but also for determining the diversity, for instance, of a molecular population of mobile genetic element lineages in a host genome. In other words, the concept of community can be considered at both ecological and molecular levels. Taking this into account, CheckAlign 2.0 computes two variations of the Shannon algorithm. For constructing sequence logos from the input of gapped and ungapped protein and DNA multiple alignments, CheckAlign 2.0 computes the canonical algorithm employed in CheckAlign 1.0, which has been described in depth ( and references therein). For accounts of both the abundance and evenness of the species present in a community sample, CheckAlign 2.0 computes the Shannon-Weaver diversity index (H) and the Equitability (EH) according to the following algorithms, respectively.Algorithm 1
Here, S is the total number of species in the community (richness) and pi is the proportion of S made up by the i-th species. Also, HMAX = ln S (equitability assumes a value between 0 and 1, where 1 indicates complete evenness).
The CheckAlign 2.0 standalone application is distributed as an installer for Windows XP/Vista/7 (32 bit and 64 bit), a self-extracting disk image for Mac OS X 10.5 or later (64 bit), and a compressed tarball archive for Linux 2.6 kernel series or later (32 bit and 64 bit).
The standalone application requires Java 6 or later. The minimum system requirements for this software are a PC with a Pentium 4 1.5 GHz or AMD Athlon XP 1500+ processor or higher with at least 1 GB of RAM.
The development of CheckAlign 2.0 has been partly supported by Grant IDI-20100007 from CDTI (Centro de Desarrollo Tecnológico Industrial) and by Torres-Quevedo Grants PTQ-09-01-00020 and PTQ-09-01-00670 from MICINN (Ministerio de Ciencia e Innovación) in Spain.
Funding to pay the Open Access publication charges for this article was provided by the University of Valencia
License and distribution
CheckAlign 2.0 is owned by Biotech Vana S.L. and is freely available as both an online server version and as a standalone application at [URL1, and URL 2]. The software and its source code are distributed under the terms of the Eclipse Public License v1.0 [URL 8] (formerly Common Public License 1.0) considered in the agreement for open source applications that you should accept during the installation of this tool.
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- Llorens C, Futami R, Vicente-Ripolles M, Moya A: The CheckAlign logo-maker application in analyses of both gapped and ungapped DNA and protein alignments. In Biotechvana Bioinformatics. Biotechvana, Valencia; 2008:SOFT: CheckAlign.
- Checkalign 2.0 Server: http://gydb.org/tools/checkalign
- Checkalign 2.0 software: https://biotechvana.com/checkalign/index
- PHP programming language: http://php.net
- ECMAScript language specification: http://www.ecma-international.org/publications/standards/Ecma-262.htm
- HMMER: http://hmmer.janelia.org
- Sun Microsystems: http://www.java.com
- Eclipse Public License - v 1.0: http://www.opensource.org/licenses/eclipse-1.0.php