3. Results
3.1. Characterization of the full-length cDNA of PmXbp1
The full-length cDNAof PmXbp1was 1762 bp in length containing an
open reading frame (ORF) of 855 bp corresponding to a polypeptide of
284 amino acidswith the 5′ and 3′ UTRs of 147 and 741 bp, respectively
(excluding the poly A tail, GenBank accession no. JN644920, Fig. 1).
PmXbp1 showed the closest sequence similarity to Xbp1 of Hottentotta
judaicus (E-value = 1e−24). The deduced PmXbp1 protein contained
three putative N-linked glycosylation sites (NXS/T) at positions
174–176, 240–242 and 280–282, respectively. The calculated pI
and MW of the deduced PmXbp1 protein were 7.65 and 30.66 kDa, respectively.
The signal peptide was not found in this presumed nonsecretory
protein. A BRLZ domain (E-value = 2.32e−10) was found at
amino acids positions 93–157 of the deduced PmXbp1 protein (Fig. 1).
3.2. Determination of sequence polymorphism in PmXbp1 using PCR-SSCP
analysis and DNA-cloning and sequencing
The PCR product of 185 bp in sizewas obtained fromamplification of
PmXbp1 against genomic DNA of 3-month-old P. monodon juveniles. A
total of 3 SSCP patterns were observed (Fig. 2). Of these, patterns A
(N = 44 accounting for 27.16%) and B (N = 116 accounting for
71.60%) were collectively observed in 98.77% of examined specimens
while pattern C was found in only two male individuals (1.23%). Only
a single SNP (T→C349)was observed fromPCR-cloning and sequencing
of representative individuals exhibiting SSCP patterns A (T/T349) and B
(T/C349) (N = 9 each).
3. Results3.1. Characterization of the full-length cDNA of PmXbp1The full-length cDNAof PmXbp1was 1762 bp in length containing anopen reading frame (ORF) of 855 bp corresponding to a polypeptide of284 amino acidswith the 5′ and 3′ UTRs of 147 and 741 bp, respectively(excluding the poly A tail, GenBank accession no. JN644920, Fig. 1).PmXbp1 showed the closest sequence similarity to Xbp1 of Hottentottajudaicus (E-value = 1e−24). The deduced PmXbp1 protein containedthree putative N-linked glycosylation sites (NXS/T) at positions174–176, 240–242 and 280–282, respectively. The calculated pIand MW of the deduced PmXbp1 protein were 7.65 and 30.66 kDa, respectively.The signal peptide was not found in this presumed nonsecretoryprotein. A BRLZ domain (E-value = 2.32e−10) was found atamino acids positions 93–157 of the deduced PmXbp1 protein (Fig. 1).3.2. Determination of sequence polymorphism in PmXbp1 using PCR-SSCPanalysis and DNA-cloning and sequencingThe PCR product of 185 bp in sizewas obtained fromamplification ofPmXbp1 against genomic DNA of 3-month-old P. monodon juveniles. Atotal of 3 SSCP patterns were observed (Fig. 2). Of these, patterns A(N = 44 accounting for 27.16%) and B (N = 116 accounting for71.60%) were collectively observed in 98.77% of examined specimenswhile pattern C was found in only two male individuals (1.23%). Onlya single SNP (T→C349)was observed fromPCR-cloning and sequencingof representative individuals exhibiting SSCP patterns A (T/T349) and B
(T/C349) (N = 9 each).
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