Evolution of Kryptolebias marmoratus from the order Teleost Using Vasa as a

molecular marker

by

Jacob L. Perry

Background

• Kryptolebias marmoratus– Relationship to - Oryzias latipes

• 1958, 1960-61- Harrington• Special feature observed

– Specialized gonad

• Temperature dependent• Special note: no known wild-type female• Used in research - recently named model

organism within last 10 years

Introduction

• Data at the present - limited in certain areas

• Vasa - DEAD-box domain, ATP-dependent-RNA helicase– Helps in the development of PGC– Cytoplasmic zone during embryogenesis

• Highly conserved region

Materials and Methods

• RNA extraction, cDNA development w/ specific primers F2/R1

• Fragment in FASTA format

• NCBI- BLAST

• Seaview-protein alignment

• RAxML7.0.4

• Figtree2.1

Figure 1. Partial alignment of Vasa DEAD-box domain

Figure 2. Phylogeney of Kryptolebias marmoratus Vasa with other Teleost.

Figure 3. Graphical map of Oryzias latipes Vasa gene.

Figure 4. Diagnostic gel of PCR samples, Vasa

Figure 5. Structure of DEAD-box domain from Drosophilia melanogaster.

Courtesy of NCBI – PDB-ID: 2DB3

References:

• Alexandros Stamatakis : ﾒ RAxML-VI-HPC: Maximum Likelihood-based Phylogenetic Analyses with Thousands of Taxa and Mixed Models ﾓ , Bioinfor- matics 22(21):2688 ﾐ 2690, 2006 [4].

• Andrew Rambaut : FigTree v1.3.1 2006-2009. Institute of Evolutionary Biology University of Edinburgha.rambaut@ed.ac.uk <http://tree.bio.ed.ac.uk/>

• Galtier, N., Gouy, M. and Gautier, C. (1996) SeaView and Phylo_win, two graphic tools for sequence alignment and molecular phylogeny. Compututer Applications in the Biosciences 12, 543–548. Available

through http://pbil.univ-lyon1.fr/software/.

• Harrington RW Jr. & Rivas LR. 1958. The discovery in Florida of the Cyprindont fish, Rivulus marmoratus, with a redescription and ecological notes. Copeia ; 2: 125-130.

• Harrington Jr., R.W. 1961. Oviparous hermaphroditic fish with internal fertilization. Science. 134: 1749-1750.

• Harrington Jr., R.W. 1963. Twenty-four hour rhythms of internal self-fertilization and of oviposition by hermaphrodites of Rivulus marmoratus. Physiological Zoology. 36: 325-341.

• Katoh, K., Misawa, K., Kuma, K., and Miyata T. (2002) MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Research 30, 3059–3066.

• Katoh, K., Kuma, K., Toh, H., and Miyata T. (2005) MAFFT version 5: improvement in accuracy of multiple sequence alignment. Nucleic Acids Research 33, 511–518. Available at http://align.bmr.kyushu-

u.ac.jp/mafft/software/.

• National Center for Biotechnology Information (NCBI) Entrez, public domain software distributed by the authors. http://www.ncbi.nlm.nih.gov/Entrez/ National Library ofM edicine, National Institutes of Health,

Bethesda, Maryland, U.S.A.