1. Evolution of Kryptolebias marmoratus from the order Teleost Using Vasa as a molecular markerby
Jacob L. Perry

2. Background Kryptolebias marmoratus Relationship to - Oryzias latipes
1958, 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

3. 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

4. RNA extraction, cDNA development w/ specific primers F2/R1
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

5. Figure 1. Partial alignment of Vasa DEAD-box domain

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

7. Figure 3. Graphical map of Oryzias latipes Vasa gene.

8. Figure 4. Diagnostic gel of PCR samples, Vasa

9. Figure 5. Structure of DEAD-box domain from Drosophilia melanogaster.
Courtesy of NCBI – PDB-ID: 2DB3

10. 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 v Institute of Evolutionary Biology University of <
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
Harrington RW Jr. & Rivas LR The discovery in Florida of the Cyprindont fish, Rivulus marmoratus, with a redescription and ecological notes. Copeia ; 2:
Harrington Jr., R.W Oviparous hermaphroditic fish with internal fertilization. Science. 134:
Harrington Jr., R.W Twenty-four hour rhythms of internal self-fertilization and of oviposition by hermaphrodites of Rivulus marmoratus. Physiological Zoology. 36:
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 u.ac.jp/mafft/software/.
National Center for Biotechnology Information (NCBI) Entrez, public domain software distributed by the authors. National Library ofM edicine, National Institutes of Health, Bethesda, Maryland, U.S.A.