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W Petchpoo P Tan-ariya V Boonsaeng C R Brockelman P Wilairat S Panyim 《Veterinary parasitology》1992,42(3-4):189-198
A genomic library of Babesia bovis DNA from the Mexican strain M was constructed in plasmid pUN121 and cloned in Escherichia coli. Several recombinants which hybridized strongly to radioactively labeled B. bovis genomic DNA in an in situ screening were selected and further analyzed for those which specifically hybridized to B. bovis DNA. It was found that pMU-B1 had the highest sensitivity, detecting 25 pg of purified B. bovis DNA, and 300 parasites in 10 microliters of whole infected blood, or 0.00025% parasitemia. pMU-B1 contained a 6.0 kb B. bovis DNA insert which did not cross-hybridize to Babesia bigemina, Trypanosoma evansi, Plasmodium falciparum, Anaplasma marginale, Boophilus microplus and cow DNA. In the Southern blot analysis of genomic DNA, pMU-B1 could differentiate between two B. bovis geographic isolates, Mexican strain M and Thai isolate TS4. Thus, the pMU-B1 probe will be useful in the diagnosis of Babesia infection in cattle and ticks, and in the differentiation of B. bovis strains. 相似文献
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In vitro assembly of Penaeus monodon densovirus (PmDNV)‐like particles produced in a prokaryote expression system 
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下载免费PDF全文 Rapee Sinnuengnong Pongsopee Attasart Duncan R Smith Sakol Panyim Wanchai Assavalapsakul 《Aquaculture Research》2017,48(9):4975-4981
Viral diseases are a significant problem in the shrimp aquaculture industry as outbreaks can cause significant mortality and economic loss. While it has been shown that triggering the shrimp RNA interference pathway through dsRNA is a potentially viable treatment pathway, this approach is hampered by the lack of a suitable delivery mechanism. Virus‐like particles (VLPs), which are structurally similar to native viruses but lack the genetic material, could possibly be developed as a delivery vehicle. To generate a candidate VLP, the Penaeus monodon densovirus (PmDNV) capsid protein was cloned with an added histidine tag and expressed in an E. coli expression system. While the protein was expressed in inclusion bodies, the recombinant PmDNV capsid protein could be dissolved and subsequently purified by nickel affinity column chromatography. The formation of VLP from this purified rPmDNV capsid protein was investigated by transmission electron microscopy, and PmDNV‐VLPs were observed that looked similar to the native PmDNV virion. Our results suggest that the PmDNV‐like particle could be promisingly applied towards vaccination and that this PmDNV‐like particle can potentially serve as a system for delivery of nucleic acids to trigger innate immunity in shrimp. 相似文献
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In vitro study of a putative role of gonad‐inhibiting hormone in oocyte growth stimulation in Penaeus monodon 下载免费PDF全文
下载免费PDF全文 Ovarian development in crustacean is controlled by several factors, among which a neuropeptide gonad‐inhibiting hormone (GIH) is known to inhibit vitellogenin (Vg) synthesis in the ovary. It has been postulated that GIH may control Vg synthesis by inhibiting the release of gonad‐stimulating factor (GSF) from brain and thoracic ganglia. To prove this hypothesis, this study was primarily aimed to investigate the influence of GIH on the release of GSF from thoracic ganglia of Penaeus monodon. Our result showed that GIH did not suppress the release of putative GSF from thoracic ganglia by calcium ionophore A23187 as the induction of oocyte growth in the ovary explants that were cocultured with thoracic ganglia in the presence of A23187 was not affected by the addition of recombinant GIH protein. In addition and interestingly, when the ovary explants were incubated with the recombinant GIH alone, the oocyte growth was increased at the rate comparable to that induced by A23187 in the presence of thoracic ganglia. Hence, our in vitro study demonstrated that the stimulation of GSF released from thoracic ganglia is independent of GIH, and that the GIH has a dual function in oocyte growth stimulation and inhibition of Vg synthesis in the early stage of ovarian development. This expands our knowledge on the regulation of ovarian development in shrimp by GIH. Further in vivo studies in this novel aspect of GIH function will be useful for the improvement of shrimp ovarian maturation in the future. 相似文献
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