Aquaculture 265 (2007) 16 – 26

www.elsevier.com/locate/aqua-online

Cloning of eIF5A from shrimp Penaeus monodon, a highly

expressed protein involved in the survival of WSSV-infected shrimp
Amornrat Phongdara b,⁎, Yanisa Laoong-u-thai a , Warapond Wanna b
a
Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand
b
Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla 90112, Thailand
Received 19 August 2006; received in revised form 22 December 2006; accepted 4 February 2007

Abstract

Shrimp respond to viral infection with up- and down-regulation of many critical genes. In our previous work, we showed that
white spot syndrome virus (WSSV) infection of Penaeus monodon (Pm) caused cellular syntenin levels to increase. In order to
further explore the signal transduction pathway of syntenin, we constructed the cDNA library of WSSV-infected shrimp and
performed a yeast two-hybrid screening of the library using syntenin as bait. Here we report that syntenin specifically binds
eukaryotic translation initiation factor 5A (eIF5A), a 157 amino acid polypeptide that is implicated in cell proliferation and
survival. GST-pull-down assays showed the presence of specific interaction between Pm-syntenin and eIF5A. Message analyses
revealed that syntenin expression remained normal when shrimp were infected with WSSV but did not show any gross signs of
disease. Syntenin expression, however, increased 1.4-fold when shrimp became dead. In contrast, eIF5A expression increased
markedly during the stage of WSSV infection when there were no gross signs of disease, i.e., prior to the moribund stage. At the
moribund stage, eIF5A expression was down-regulated. This data suggest that syntenin and eIF5A physically interact with each
other in the setting of WSSV infection and that these two molecules exhibit dynamic changes in their expression patterns as
infection progresses from grossly normal, infected shrimp to full-blown phase.
© 2007 Elsevier B.V. All rights reserved.

Keywords: eIF5A; Translation initiation; Syntenin; Protein–protein interaction; Yeast two hybrid

1. Introduction shrimp, the white spot syndrome virus (WSSV) is

Penaeid shrimp production is a worldwide economic
activity. The intensification of shrimp farming over the
last few decades has been accompanied by develop-
ment of infectious diseases of viral, bacterial, and, in
some cases, fungal origin (Destoumieux-Grazon et al.,
2001). Among the various viruses of the penaeid
⁎ Corresponding author.
E-mail address: pamornra@yahoo.com (A. Phongdara).
0044-8486/$ - see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.aquaculture.2007.02.005
responsible for a major proportion of the diseases
plaguing commercial shrimp ponds, and has resulted in
high mortality and economic losses. (Lightner, 1996;
Flegel, 1997, 2001).
Shrimp exhibit a diverse response to viral infection,
that is manifested in marked up- and down-regulation of
a variety of genes (Gross et al., 2001; Supungul et al.,
2002; Rojtinnakorn et al., 2002; Sotelo-Mundo et al.,
2003; Hikima et al., 2003; He et al., 2005; De Lorgeril
et al., 2005; Du et al., 2006). In our previous work, we
identified the protein syntenin of the shrimp Penaeus
 A. Phongdara et al. / Aquaculture 265 (2007) 16–26
monodon (Pm) as a dynamic responder to white spot
syndrome virus (WSSV) infection, its message being
greatly up-regulated in the acute phase of the infection
(Bangrak et al., 2002). Reports from studies of other
organisms indicate that syntenin is involved in the signal
transduction pathway. Syntenin has no obviously
catalytic domain and therefore is unlikely to have a
signaling function by itself, but could serve as an
adapter or scaffolding protein to attach other proteins
and functions in the cell. As of now several syntenin-
binding proteins from other organisms have been
identified and their functions have been described.
These include syndecan (Grootjans et al., 1997), pro-
TGFα (Femandez-Larrea et al., 1999), ephrin B (Torres
et al., 1998; Lin et al., 1999), neurofascin (Grootjans
et al., 2000), interleukin 5 receptor α subunit (IL5Rα),
and EphA7 (Torres et al., 1998).
In order to further explore the link between Pm-
syntenin and viral infection, we performed a yeast two-
hybrid screening of a P. monodon cDNA library, using
Pm-syntenin as bait. This experiment resulted in seven
independent clones of syntenin-binding proteins being
isolated. Among these syntenin-binding proteins, one
(α2M) that plays an important role in immune mechan-
isms in shrimp has already been described (Tonganunt
et al., 2005). Here we describe the identification and
characterization of another syntenin-binding protein
eIF5A and we follow its expression during the course
of a WSSV infection.
2. Materials and methods
2.1. Samples
All shrimp were obtained from a farm controlled by
the Songkhla Coastal Fisheries Research and Develop-
ment Center, Thailand. The shrimp were sourced from a
pond stocked with postlavae that had tested negative for
WSSV by nested PCR (Lo et al., 1996a,b) and that later,
when the pond was harvested, showed no sign of WSSV
infection. Prior to the beginning of the experiment,
shrimp were kept individually in 60 l aquaria for
10 days, all samples were checked for WSSV infection
by using PCR technology (Lo et al., 1996a,b).
Hemolymph fluid (100–150 μl) was initially withdrawn
and labeled as normal (NS) for individual shrimp whose
PCR results proved negative for WSSV. Hemolymph
from individual shrimp identified by PCR as WSSV
positive, but showing no sign of the white spot
syndrome, was labeled as originating from grossly
normal, infected shrimp (GNIS). WSSV experimental
infections were carried out by injection of 10 μl of a
17
1:1 × 107 dilution of a viral stock solution made in
0.85% NaCl and 100 to 150 μl of hemolymph was
subsequently withdrawn at 6, 12, 24, 48 and 72 h
postinjection (p.i.). The remaining shrimp were held up
to the time when they became moribund (shrimp is
immobile and unresponsive to touch but were still
showing movement of gill rakers. At this time it was
labeled as moribund, infected shrimp (MIS). When gill
movement ceased, hemolymph was immediately with-
drawn and labeled as dead shrimp (DIS). Hemolymph
was stored at − 80 °C until used.
2.2. Plasmid construction
Proteins expressed using vector pGBKT7, were
fused with amino acids 1–147 of the GAL4 DNA
binding domain (BD). Proteins expressed using vector
pGADT7 were fused with amino acids 768–881 of the
GAL4 activation domain (AD). The control plasmids
pGBKT7-53 and pGADT7-T were obtained from
CLONTECH Laboratories, Inc. The Plasmid BD-
syntenin containing a full-length Pm-syntenin gene
was constructed by inserting the syntenin DNA in-
frame into an EcoRI and BamHI site. Plasmid
construction was verified by sequencing cDNA from
the hemocytes of WSSV-infected shrimp, ligated into the
AD-prey vector that had been previously digested with
the EcoRI and XhoI as reported from previous work
(Tonganunt et al., 2005).
2.3. Yeast two-hybrid screening
Yeast two-hybrid screening was carried out with the
MATCHMAKER Gal4 Two-hybrid System 3 (CLON-
TECH Laboratories, Inc.). Competent cells of the yeast
strain AH109 were prepared and transformations were
performed according to the manufacturer's protocols.
Briefly, 1 μg of plasmid DNA was added to 100 μl of
competent cells and mixed again with 36 μl of 1 M
lithium acetate and 240 μl of 50% polyethylene glycol
(PEG). The transformation mixture was then incubated
at 30 °C for 30 min followed by a heat-shock at 42 °C
for 15 min and subsequently spread on a drop-out-agar
plate in the absence of leucine and tryptophan. The
plates were incubated at 30 °C for 48 h to allow for yeast
growth. PCR was used to confirm transformation with
the target gene. A positive clone was inoculated into SD
medium lacking leucine, tryptophan and histidine (SD-
LTH) and supplemented with 3-amino-1,2,4-triazole
(SD-LTH + 5 mM 3-AT). The medium was shaken at
180 rpm at 30 °C for 96 h. Absorbance of the medium at
595 nm was measured in a spectrophotometer
18 A. Phongdara et al. / Aquaculture 265 (2007) 16–26

method used here is reproducible and of equal or greater

sensitivity in comparison with the β-galactosidase assay
(Diaz-Camino et al., 2003). All of the yeast media used
were prepared according to the standard protocols
(Handbook PT3024-1, CLONTECH Laboratories, Inc.).

2.4. Sequence analysis

Plasmid DNA was isolated from the clones con-

Fig. 1. Growth analysis results for the yeast two-hybrid assay. S.
cerevisiae AH109 cell were cotransformed with BD-syntenin and AD-
sbp (marked as Syn + Sbp). Transformed cells were inoculated in SD
medium. For the positive control yeast cells were transformed with
pGBKT7-53 and pGADT7-T (positive) and the empty host cells were
used as the negative control (negative).
(PerkinElmer instruments). This is an alternative
method of screening colonies for transformants based
on the growth curve analysis for yeast culture. The
structed in Section 2.2 and transformed into Escherichia
coli Top10F′ for plasmid recovery and sequencing. The
sequence of the eIF5A gene of P. monodon was
deposited at GenBank under accession number
DQ851145. The searches for nucleotide and protein
sequence similarities were conducted with BLAST
algorithm at the National Center for Biotechnology
Information (NCBI) (http://www.ncbi.nlm.nih.gov/
BLAST/). The eIF5A deduced amino acid sequence
was analyzed with the NCBI Protein Sequence Analysis
software. Multiple alignment of the eIF5A was per-
formed with the ClustalX Multiple Alignment program
(http://www.ncbi.nlm.nih.gov) and Multiple Alignment

Fig. 2. Nucleotide and deduced amino acid sequences of P. monodon eIF5A cDNA. Amino acids are indicated as single capital letters under each
triplet codon of the nucleotide sequence. The 12-amino acid hypusine core region is shown in bold type while the hypusine modification site is shown
in bold underlined type. An asterisk (⁎) indicates the stop codon. The polyadenylation signal (ATTAAA) is underlined.
 A. Phongdara et al. / Aquaculture 265 (2007) 16–26 19

Fig. 3. Phylogenetic analysis of the eIF5A amino acid sequences of P. monodon and various other species: S. frugiperda, S. exigua, H. sapiens, B.
mori, D. rerio, Rabbit, A. mellifera, C. familiaris, M. musculus, R. norvegicus, X. laevis, B. taurus, G. gallus, P. troglodytes, P. pygmaeus, X.
tropicalis, and D. melanogaster obtained from GenBank. B. taurus was used as the out-group. Analysis was based on the completed amino acid
sequences.

show by GENEDOC, version 2.6.001 (Nicholas and
Nicholas, 1997).
2.5. Expression and purification of recombinant
Pm-syntenin
A BamHI-PstI fragment containing the entire coding
region of Pm-syntenin was obtained from previous
work (Bangrak et al., 2002). This recombinant plasmid
as well as an insertless pQE40 expression vector were
transformed into E. coli M15 and proteins were prepared
according to Bangrak et al. (2002).
pGEX-4T-1 expression vector (Amersham Biosciences)
that had been digested with the same restriction
enzymes. The resultant plasmid pGEX4T-1-eIF5A was
transformed into E. coli BL21 (Amersham Biosciences).
The cell was cultivated in medium containing 100 μg/ml
of ampicillin and the expression of a recombinant
protein was induced by the addition of 0.1 mM IPTG.
Cells were harvested and the GST-eIF5A fusion protein
was purified by using a Glutathione–Sepharose 4B resin
column (Amersham Biosciences). The purified protein
was detected as a single band on a 12% SDS-PAGE, and
kept at − 80 °C until ready for use.

2.6. Expression and purification of GST-tagged eIF5A
protein
The AD-plasmid harboring the eIF5A gene was
digested with EcoRI and XhoI, then ligated into the
2.7. In vitro binding assays (in vitro GST-pull-down assays)
GST-pull-down assay was performed using the Bulk
GST purification Module (Amersham Biosciences). The
purified GST-eIF5A fusion protein was adsorbed onto
20 A. Phongdara et al. / Aquaculture 265 (2007) 16–26

Fig. 4. In vitro binding assay. A glutathione–sepharose bead pull-down was performed on the combined proteins as shown in the table (nos. 1, 3, 4, 6).
The eluted material was loaded on SDS-PAGE gels, transferred then detected using specific antibodies. A GST band and GST-eIF5A were detected
with anti-GST antibody (lanes 4–6). The eluted material was also detected for syntenin with anti-His Tag antibody (lanes 1, 3). Lane 2 and lane 5 are
purified 6xHis-syntenin and GST-eIF5A, used as a control for antibody detection.

Glutathione–Sepharose beads (20 μl of a 50% bed
slurry) for 1 h, then washed ten times and finally
resuspended in phosphate-buffered saline, pH7. The
beads carrying the GST-eIF5A fusion protein would
serve as the bait protein in the subsequent steps. The
prey protein, a purified 6xHistidine-tagged protein fused
with syntenin (6xHis-syntenin) was combined with the
bait protein and the mixture was gently shaken for 2 h at
room temperature (RT). The proteins in solution were
washed 6 times using 100 μl of phosphate-buffered
saline and eluted by the addition of a buffer containing
reduced glutathione. The eluted samples were analyzed
by 12% SDS-PAGE and transferred onto a nitrocellu-
lose membrane. The blots were incubated with an anti-
His Tag antibody (His-Probe horseradish peroxidase
conjugated, Pierce; diluted 1:20,000) and visualized
using ECL detection reagent (Pierce). To confirm the
presence of the GST-fusion protein, the blots were
incubated with Anti-GST (Amersham Biosciences;
1:2000) and conjugated goat anti-mouse IgG-alkaline
phosphatase (Pierce; diluted 1:20,000). Lumi-Phos was
used as a substrate for detecting chemiluminescence.
2.8. Semi-quantitative RT-PCR
Total cellular RNA of P. monodon hemocytes was
isolated from NS (uninfected shrimp), GNIS and DIS
using TRIZOL Reagent (Life Technologies). The total
RNA (800 ng) from each sample was reverse tran-
scribed to produce cDNA using SuperScript™ III Re-
verse Transcriptase (Invitrogen). The resulting cDNA
(500 ng) was amplified by Taq DNA polymerase
(Promega) and 0.2 μM of primers specific for each
gene. The following thermal profile was used for PCR
amplification of the cDNA on a GeneAmp® PCR
system 9700 (Applied Biosystems): 1 cycle of 94 °C for
5 min followed by 25 cycles of 94 °C for 1 min, 50 °C
for 1 min and 72 °C for 1 min, with a final extension at
72 °C for 10 min. PCR products were electrophoret-
ically separated on a 1.5% agarose gel, stained with
ethidium bromide and visualized under ultraviolet light
by Gel Doc 1000 genetic Analyzer (BIO RAD). Semi-
quantitation by RT-PCR was carried out in triplicate.
2.9. SYBR Green RT-PCR
cDNA was synthesized in a 10 μl reaction volume
containing 800 ng of DNase I treated total RNA and
SuperScript™ III Reverse Transcriptase (Invitrogen).
For comparing the detection limits of the amplicon in
SYBR Green PCR, 300 ng cDNA was used per reaction.
The reaction mixture contained 12.5 μl of iQ™ SYBR®
Green Super Mix (BIO RAD) and 0.2 μM of each
forward and reverse primer. The thermal profile for the
PCR amplification was 95 °C for 10 min, followed by
40 cycles of 94 °C for 1 min, 50 °C for 1 min and 72 °C
for 1 min. A post-PCR curve was run according to the
MX3000P™ (STRATAGENE®): a 1 min hold at 95 °C,
a 30 s hold at 50 °C, and a 30 min slow ramp from 50 to
95 °C. Each sample was comprised of 3 replicates and
all reactions were independently repeated twice to
ensure the reproducibility of the results. For real-time
 A. Phongdara et al. / Aquaculture 265 (2007) 16–26 21

Fig. 5. (A) RT-PCR assay of eIF5A, syntenin and α2M gene expression in hemocytes of 4 individuals of uninfected P. monodon (NS), grossly
normal, infected shrimp (GNIS) and dead shrimp (DIS). The expression is compared to elongation factor (EF) as an internal control. (B) Normalized
eIF5A, syntenin, α2M and eIF4A expression were calculated from the images using Scion Images software. The bars represent means with standard
deviation (SD) for results obtained from 4 individuals in each sample set.

RT-PCR quantification of eIF5A and syntenin expres-
sion, the shrimp gene elongation factor-1α (EF) (Dhar
et al., 2002) was used as an internal control. Anova
comparison tests were used for statistical analysis by
SPSS software (version 14.0). Values were considered
to be significant at P b 0.05.
3. Results
3.1. Identification of Pm-syntenin-binding protein
Using the yeast two-hybrid method with Pm-
syntenin as the bait and a WSSV-infected shrimp
hemocyte library as the prey, we identified 7 yeast
clones that contained possible syntenin-binding proteins
(Sbps) as a result of their growth in high stringency
medium (SD-LTH + 5.0 mM 3-AT). An example of the
growth analysis is shown in Fig. 1 and includes the
clone showing the best growth (Syn + Sbp). By contrast,
empty host control cells gave no growth (Fig. 1). The
prey vector from this clone was extracted and
transformed into E. coli for plasmid recovery and
sequencing.
3.2. Characterization and comparison of syntenin-
binding protein (Sbp) sequences
Sequencing of the cDNA insert in the plasmid
vector of the fastest growing yeast clone revealed a
deduced protein sequence of 157 amino acids with a
predicted molecular mass of 17.27 kDa (Fig. 2). The
deduced sequence showed 81% identity to the protein
eIF5A of the moth Spodoptera frugiperda (GenBank
accession number AAF13316.1). Alignment of the
P. monodon protein sequence with other eIF5A se-
quences at GenBank revealed that eIF5A is a highly
conserved protein, particularly at the N-terminal por-
tion. In addition to the complete conservation of 12
amino acids constituting the hypusine core region
(bold letters in Fig. 2), several other amino acids were
also conserved. These included 5 proline residues,
indicating conservation of the secondary structure of
the protein. Phylogenetic analysis of eIF5A amino acid
sequences of P. monodon and eIF5As from various
other species showed that the eIF5A of P. monodon
fell within a group of arthropod (insect) proteins
(Fig. 3).
22 A. Phongdara et al. / Aquaculture 265 (2007) 16–26

Fig. 6. Real-time PCR machine output and standard curves for eIF5A, syntenin and α2M.

3.3. Interaction of syntenin with eIF5A in vitro 3.4. Gene expression by semi-quantitative RT-PCR

Pull-down assays using soluble His-labeled Pm-
syntenin with GST-labeled eIF5A bound to sepharose
beads confirmed binding between the two proteins
(Fig. 4). Specifically, western blot detection of His and
GST labels revealed that both were present in the eluant
(Fig. 4, lanes 3 and 6) from glutathione treated beads
after exposure to His-labeled syntenin. However, no His-
labeled syntenin was found when GST alone was bound
to the beads (Fig. 4, lanes 1 and 4). This test confirmed
that syntenin binds specifically to eIF5A and not to GST.
Semi-quantitative RT-PCR during the course of a
WSSV infection revealed that the expression of eIF5A
was high in grossly normal WSSV-infected shrimp
(GNIS) when compared to normal shrimp (NS) and
dead WSSV-infected shrimp (DIS) (Fig. 5). Since α2M is
also known as a syntenin-binding protein, we also
measured its expression during the course of WSSV
infection and found that it was also high in GNIS when
compared to NS and DIS. In addition, we measured the
expression of eIF4A, a member of the initiation factor
 A. Phongdara et al. / Aquaculture 265 (2007) 16–26 23

Fig. 7. eIF5A, syntenin and α2M gene expression by quantitative real-time RT-PCR. (A) Fold-change (x-change, mean ± S.D.) of eIF5A and
syntenin mRNA expression in hemocytes of 3 individuals of uninfected P. monodon (normal, NS), grossly normal, infected shrimp (GNIS)
and dead shrimp (DIS). The expression level was compared to elongation factor as an internal control. The standard curve method was used
for the quantification of mRNA, NC = normal control. (B) Fold-change (x-change, mean ± S.D.) of eIF5A and syntenin mRNA expression in
hemocytes of experimentally infected shrimp at various times post challenge (6, 12, 24, 48, 72 h postinjection; MIS = moribund infected
shrimp; DIS = dead shrimp).

complex that controls protein translation (Palacios et al.,
2004), and we found that its expression differed
markedly from that of eIF5A (Fig. 5). Expression was
relatively low for all samples.
3.5. Syntenin and eIF5A gene expression by quantita-
tive RT-PCR assay
Real-time PCR amplification curves for the three
genes generated by the MX3000P™ (STRATAGENE®)
were very reproducible and indicated that the primers
were selective and effective in producing the specific
PCR products (Fig. 6A,C,E). The accuracy of mRNA
quantification, and sensitivity and linearity of SYBR
Green-based Q-RT-PCR were examined using a 10-fold
serial dilution of each DNA standard. The relationship
between threshold cycle (CT) and the log copy number
of the DNA standard was linear with r2 N 0.97 for the
three genes, indicating that the CT values changed
proportionally with serial dilution of the samples. The
reproducibility of the techniques within and between
assays was tested, using serial dilutions of eIF5A,
syntenin and α2M cDNA standards.
By reference to the standard curves, expression of
eIF5A and α2M was high in GNIS but low in DIS when
compared to NS (Fig. 7A). The increased expression in
GNIS when compared to NS was 2.7-fold for eIF5A and
4.6-fold for α2M. However, both then decreased by 3.2-
fold and 4.7-fold, respectively in DIS. By contrast,
syntenin expression appeared to increase gradually as the
disease progressed, reaching 1.4-fold that of NS in DIS.
A more detailed time-course examination of eIF5A
and syntenin expression during the course of WSSV
infection (Fig. 7B), revealed general trends for eIF5A
expression. An initial rise was followed by a decline as
the shrimp approached death, while syntenin expression
fell initially and finally rose sharply as the shrimp
approached death.
24 A. Phongdara et al. / Aquaculture 265 (2007) 16–26
4. Discussion
The yeast two-hybrid assay is a powerful method for
identifying protein–protein interactions that we have
previously used to successfully identify several proteins
binding to known bait proteins (Tonganunt et al., 2005;
Senapin and Phongdara, 2006). After protein identifi-
cation, we can further investigate and characterize the
function of these binding proteins. Here, we have
shown that eIF5A derived from a cDNA library of
hemocytes from WSSV-infected shrimp can specifically
bind with syntenin, a suggested adapter protein
previously identified in shrimp hemocytes. Although
eIF5A binding with syntenin has been previously
reported by Li et al. (2004), ours is the first report of
the phenomenon from the cDNA library of a marine
animal. We earlier reported that syntenin was up-
regulated at the acute stage of WSSV infections in
shrimp (Bangrak et al., 2002) and similar results were
obtained again here using both semi-quantitative and
quantitative RT-PCR analysis. The initial increase in
eIF5A expression during the period of infection, when
no gross signs of disease are visible, indicates that
eIF5A may play some role in slowing disease prog-
ression. This is supported by the fact that its expression
falls as the shrimp approach death. It has been sug-
gested (Li et al., 2004) that binding of syntenin and
eIF5A may act to prevent apoptosis and apoptosis has
been proposed as a cause of death in WSSV-infected
shrimp (Flegel, 2006). Thus, we may hypothesize that
eIF5A binding to syntenin prevents apoptosis during the
early stages of WSSV infection but fails to do so as the
expression of eIF5A declines with disease progression.
Although the function of α2M has been discussed in
our earlier work (Chotigeat et al., 2006), the function of
eIF5A remains an open question. The results from our
work and elsewhere indicates that it is responsible for
supporting viral replication while keeping the cell viable.
Recent research has revealed that eIF5A is not an
initiator of protein translation (Kang and Hershey, 1994;
Zuk and Jacobson, 1998) although it could be involved
in the translation of a specific subset of mRNAs (Park et
al. 1997). It has also been correlated with cell
proliferation (Kang and Hershey, 1994). Li et al.
(2004) demonstrated the binding of eIF5A and syntenin
and proposed a new biological activity for eIF5A as a
regulator of p53. Their data showed that the eIF5A
interaction with syntenin prevented it from inducing
increased expression of p53 protein. In addition, eIF5A
expression itself is significantly increased during
dendritic cell maturation, and hypusine formation
appears to be essential for this process (Kruse et al.,
2000). Ling et al. reported that eIF5A was involved in the
apoptosis of tumor cells induced by inhibition of
ubiquitin proteasomes. These reports indicate that
eIF5A may be involved in cell growth and apoptosis.
The finding that eIF5A is a cellular cofactor of HIV-1
Rev and HTLV-I Rex transactivator proteins indicates
that eIF5A functions as part of or provides access to a
cellular nuclear-cytoplasmic RNA transport system and
therefore supports viral replication (Bevec et al., 1996;
Junker et al., 1996; Ruhl et al., 1993; Katahira et al.,
1995; Schatz et al., 1998; Elfgang et al., 1999). Van Oers
et al. (1999) reported that the S. frugiperda genome had
a single copy of an eIF5A gene but that it was
transcribed into 4 different transcripts. Infection of S.
frugiperda cells with a baculovirus resulted in a strong
decrease in the number of all four transcripts as soon as
12 h after infection. They suggested that eIF5A was an
essential protein in insect species and that depletion of
this factor towards the end of a baculovirus infection
was likely to reduce cell viability.
A perplexing aspect of the eIF5A protein is that high
expression occurs only in grossly normal, infected
shrimp (GNIS) but not in uninfected normal shrimp
(NS) or in moribund/dead shrimp. Perhaps this is a
result of the cell's attempt to survive viral infection. It is
expected that future research will result in the
identification of more components of both the viral
and host genomes and that this will allow us to better
understand how host cells survive viral infection.
Acknowledgements
This work was supported by National Research
Council of Thailand, The Royal Golden Jubilee
Graduate Program from the Thailand Research Fund
to Ms. Yanisa Laoong-u-thai (PHD/0250/2546). cDNA
library was constructed from the project: Genomic
Researches for Increasing Culture Efficiency of the
Black Tiger Shrimp (Penaeus monodon) Phase I and
Phase II: (BT-B-06-SG-09-4603) supported by
NSTDA, Thailand. We thank Prof. Dr. Kenichi Fujise,
University of Texas Health Science Center at Houston,
and Prof. Dr. Brian Hodgson, Prince of Songkla
University for reading the manuscript and making
valuable comments.
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