Identification of three protein disulfide isomerase members from Haemaphysalis longicornis tick

Functional analysis of protein disulfide isomerases in blood feeding, viability and oocyte development in Haemaphysalis longicornis ticks

“…The data are expressed as the ratio of the density of Vg to the density of the actin gene products from the same template. nymphal and adult stages and primarily expressed in the ovary of female adult ticks (Liao et al, 2007a). The disruption of HlPDI-1 in this study caused the oocytes in the ovary to remain immature and the laying of only a few eggs, further confirming that HlPDI-1 is involved in ovarian and egg development.…”

“…Approximately 1000 bp fragments of HlPDI-1, HlPDI-2 and HlPDI-3 genes were amplified by PCR from three recombinant plasmids, which contained the full-length sequence of each mature HlPDI (Liao et al, 2007a), with oligonucleotide T7 forward and T7 reverse primers (Table 1) to attach the T7 promoter recognition sites on both 5 0 and 3 0 termini. The PCR products were purified using a gel purification kit (GENECLEAN, Bio 101 System).…”

“…The RNA samples were then used for real-time PCR and RT-PCR. Real-time PCR was performed as described previously (Liao et al, 2007a). The RT-PCR analysis was performed using a one-step RNA PCR kit (Takara, Japan) according to the instructions given with the primer sets for dsRNA construction without a T7 promoter (Table 1).…”

“…The blots of the partially fed female tick proteins (2.5 mg/lane) were incubated with antiserum against HlPDI-1, HlPDI-2 and HlPDI-3, respectively (Liao et al, 2007a). The blots of hemolymph proteins (1 mg/lane) were incubated with antiserum against Vg (Boldbaatal et al, unpublished data).…”

“…In our previous study, three genes encoding putative PDIs were identified from H. longicornis, and designated as HlPDI-1, HlPDI-2, and HlPDI-3 (Liao et al, 2007a). All three PDI genes contain two typical PDI active sites and encode putative 435, 499 and 488 amino acids, respectively.…”