FORTILIN, A SHRIMP VIRUS-RESISTANCE GENE THAT IS ALSO IMPLICATED IN THE HUMAN ACUTE ALLERGIC RESPONSE, CONTAINS RETROTRANSPOSONS-LIKE REPEATS AND IS POLYMORPHIC IN CULTURED AND WILD SHRIMP
Acacia Alcivar-Warren1-2, Natalie Isaksson1-2*, Alexandra Bowen2; Jacqueline Presedo2, Laura Hake2
1Environmental Genomics, Inc., 6 Sunrise Drive, Southborough MA 01772 USA
2Boston College, 140 Commonwealth Avenue, Chestnut Hill, MA 02467; E-mail: environmentalgenomics.warren@gmail.com
1Environmental Genomics, Inc., 6 Sunrise Drive, Southborough MA 01772 USA
2Boston College, 140 Commonwealth Avenue, Chestnut Hill, MA 02467; E-mail: environmentalgenomics.warren@gmail.com
As part of our efforts to develop a hypoallergenic shrimp line, we search for polymorphisms in allergy-causing genes across species.1 Fortilin, also known as Translationally Controlled Tumor Protein (TCTP), p23, and IgE-dependent histamine-releasing factor, is a highly conserved protein that plays a role in early heat-stress adaptation, apoptotic function, and acute allergic response.2 Fortilin binds Ca2+ and blocks Ca2+-dependent apoptosis in vivo.3 It is a growth-, cell cycle- and glucose-regulated protein.4 The complete genomic organization of a shrimp Fortilin gene is available for Marsupenaeus japonicus5 (Genbank EU294260, 4683 nucleotides). P. monodon fortilin has anti-apoptotic properties and is present at high levels during the onset of viral infections.6 Injection of rFortilin protected against White Spot Syndrome Virus (WSSV) infection resulting in 80-100% survival and detection of very low levels of WSSV by PCR, whereas in moribund samples WSSV levels were very high. Considering that the major shrimp allergens are proteins that control calcium flow in the muscular sarcoplasm, calcium levels are highly variable in shrimp, and WSSV-like ORF are present in the shrimp genome7, it is possible that transposable elements (TEs) also influence the expression of calcium-binding allergens. The aims of this study were to 1) search for TE-like repeats in Fortilin (EU294260) and compare them to those found in Fortilin sequences isolated from genomic and cDNA libraries of penaeids, and 2) develop EST-SSR markers suitable for linkage mapping. Blastn searches identified 11 Fortilin mRNAs in Genbank (4 P. monodon, 2 M. japonicus, 2Fenneropenaeus chinensis, 1 F. merguiensis, 1 L. vannamei, 1 F. indicus), in addition to 251 ESTs from healthy and YHV-, WSSV-, TSV- and Vibrio harveyi-challenged shrimp.
Various TE-like repeats were identified in Fortilin EU294260 using CENSOR software (Table 1). There are 214 Fortilin ESTs in the L. vannamei Unigene database (Lva.741) and it is expressed in most tissue types (gills; lymphoid organ; hemocytes; nerve cord; hepatopancreas; eyestalk, hemolymph). Fortilin sequences varied among and within shrimp species. Bioinformatics analyses revealed that the target site duplication sequences of long terminal repeat (LTR) retrotransposons (AGTT, CGTC), the transposon-mediated single locus sequence tag (GTCT), and the GAGGAG ribosomal binding site of pseudogenes are present in Fortilin cDNAs. Using the TEclass software, various classes of TE-like repeats were found in the 214 ESTs including 7 DNA transposons, 47 LTR retrotransposons, 64 LINE non-LTR retrotransposons, 2 SINEs and 94 unable to classify.
Polymorphic EST-SSRs were developed using primers designed from two EST sequences isolated from a cDNA library of TSV-challenged SPF L. vannamei (Fig 1). Preliminary informative alleles will be presented showing variation in wild and cultured shrimp, between SPF shrimp lines, and among reference and resource mapping families.
References
1Presedo et al. 2011. IMSEGI Session, Aquaculture America 2011, New Orleans, LA USA, Feb 28-March 3, 2011. Abst. 492. 2Choi et al. 2009.. PLoS ONE 4(6): e5732. 3Graidist et al. 2007. Biochem. J. 408(2):181-91. 4Diraison et al. 2011. Diabetologia 54(2):368-79. 5Chen et al. 2009. Biol Rep. 36(5):1135-40. 6Tonganunt et al. 2008. Fish Shellfish Immunol. 25(5):633-7. 7Plocienniczak et al. 2011. IMSEGI Session, Aquaculture America 2011, New Orleans, LA USA, Feb 28-March 3, 2011. Abs. 497.
Fuente: https://www.was.org/WASMeetings/Meetings/ShowAbstract.aspx?Id=25639
Various TE-like repeats were identified in Fortilin EU294260 using CENSOR software (Table 1). There are 214 Fortilin ESTs in the L. vannamei Unigene database (Lva.741) and it is expressed in most tissue types (gills; lymphoid organ; hemocytes; nerve cord; hepatopancreas; eyestalk, hemolymph). Fortilin sequences varied among and within shrimp species. Bioinformatics analyses revealed that the target site duplication sequences of long terminal repeat (LTR) retrotransposons (AGTT, CGTC), the transposon-mediated single locus sequence tag (GTCT), and the GAGGAG ribosomal binding site of pseudogenes are present in Fortilin cDNAs. Using the TEclass software, various classes of TE-like repeats were found in the 214 ESTs including 7 DNA transposons, 47 LTR retrotransposons, 64 LINE non-LTR retrotransposons, 2 SINEs and 94 unable to classify.
Polymorphic EST-SSRs were developed using primers designed from two EST sequences isolated from a cDNA library of TSV-challenged SPF L. vannamei (Fig 1). Preliminary informative alleles will be presented showing variation in wild and cultured shrimp, between SPF shrimp lines, and among reference and resource mapping families.
References
1Presedo et al. 2011. IMSEGI Session, Aquaculture America 2011, New Orleans, LA USA, Feb 28-March 3, 2011. Abst. 492. 2Choi et al. 2009.. PLoS ONE 4(6): e5732. 3Graidist et al. 2007. Biochem. J. 408(2):181-91. 4Diraison et al. 2011. Diabetologia 54(2):368-79. 5Chen et al. 2009. Biol Rep. 36(5):1135-40. 6Tonganunt et al. 2008. Fish Shellfish Immunol. 25(5):633-7. 7Plocienniczak et al. 2011. IMSEGI Session, Aquaculture America 2011, New Orleans, LA USA, Feb 28-March 3, 2011. Abs. 497.
Fuente: https://www.was.org/WASMeetings/Meetings/ShowAbstract.aspx?Id=25639
