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Cao X.,Sichuan Agricultural University | Cheng A.,Sichuan Agricultural University | Cheng A.,Key Laboratory of Animal Diseases and Human Health of Sichuan Province | Wang M.,Sichuan Agricultural University | Wang M.,Key Laboratory of Animal Diseases and Human Health of Sichuan Province
Proceedings - 4th International Conference on Computational and Information Sciences, ICCIS 2012 | Year: 2012

This report showed some physico-chemical properties and structural features about UL41 protein predicted by some software and on-line tools. The analysis showed both the signal peptide and the trans membrance region are not found. Meanwhile, the protein has twenty-four potential phosphorylation sites. In addition, the protein has more hydrophilic amine acid districts than hydrophobic districts and subcellular localization mainly located at the cytoplasm with 56.5%. The prediction of secondary structure showed that random coils dominate among secondary structure elements followed by alpha helix and extended strand. The prediction of tertiary structure showed the structure of the UL41 protein had a certain level of similarity to the structure-specific nuclease FEN-1(flap endonuclease-1). The study will be a basis for the further functional study of the UL41 protein. © 2012 IEEE.


Cao X.,Sichuan Agricultural University | Cheng A.,Sichuan Agricultural University | Cheng A.,Key Laboratory of Animal Diseases and Human Health of Sichuan Province | Wang M.,Sichuan Agricultural University | Wang M.,Key Laboratory of Animal Diseases and Human Health of Sichuan Province
Proceedings - 4th International Conference on Computational and Information Sciences, ICCIS 2012 | Year: 2012

The Duck Plague Virus (DPV) is the causative agent of Duck plague, which causes significant economic losses in the commercial duck industry all over the world. Our laboratory had identified and sequenced the DPV UL41 gene. Bioinformatics analysis of DPV UL41 gene were performed by some software and online tools. The results showed that GC content of duck plague virus UL41 gene was 42.42%, which encoded a protein with 498 amino acids in the peptide. The UL41 gene sequence didn't contain a rare codons string with more than two rare codons. The construction of phylogenetic tree suggested DPV had a close evolutionary relationship with the branch of the mardivirus genus, including MeHV-1, GaHV-2 and GaHV-3. © 2012 IEEE.


Shen A.-M.,Sichuan Agricultural University | Ma G.-P.,China Rural Technology Development Center | Cheng A.-C.,Sichuan Agricultural University | Cheng A.-C.,Key Laboratory of Animal Diseases and Human Health of Sichuan Province | And 13 more authors.
Virology Journal | Year: 2010

Abstract. Background. Some UL45 gene function of Herpesvirus was reported. While there was no any report of the duck enteritis virus (DEV) UL45 protein as yet. Results. The UL45 gene and des-transmembrane domain of UL45 (named UL45 gene, 295-675bp of UL45) of DEV were amplified by PCR and subcloned into the prokaryotic expression vector pET-32a(+). The constructed recombinant plasmids were transformed into the host strain BL21(DE3) PLysS and induced by IPTG. SDS-PAGE analysis showed the UL45 gene couldn't express while UL45 gene was highly expressed. His Purify Kit or salting-out could purify the protein effectively. Using the purified protein to immunize New-Zealand rabbits and produce polyclonal antibody. The agar diffusion reaction showed the titer of antibody was 1:32. Western blot analysis indicated the purified rabbit anti-UL45 IgG had a high level of specificity and the UL45 gene was a part of DEV genome. The transcription phase study of UL45 gene showed that expression of UL45 mRNA was at a low level from 0 to 18 h post-infection (pi), then accumulated quickly at 24 h pi and peaked at 42 h pi. It can be detected till 72 h pi. Besides, western blot analysis of purified virion and different viral ingredients showed that the UL45 protein resided in the purified virion and the viral envelope. Conclusions. The rabbit anti-UL45 IgG was produced successfully and it can serve as a good tool for penetrating studies of the function of DEV UL45 protein. The transcription phase and protein characteristics analysis indicated that DEV UL45 gene was a late gene and UL45 protein may be a viral envelope protein. © 2010 Shen et al; licensee BioMed Central Ltd.


Chang H.,Sichuan Agricultural University | Cheng A.,Sichuan Agricultural University | Cheng A.,Key Laboratory of Animal Diseases and Human Health of Sichuan Province | Wang M.,Sichuan Agricultural University | And 10 more authors.
Virology Journal | Year: 2010

Background. The gE protein of duck plague virus is the important membrane glycoprotein, its protein characterization has not been reported. In this study, we expressed and presented the characterization of the DPV gE product. Results. According to the sequence of the gE gene, a pair of primers were designed, and the DNA product with 1490bp in size was amplified by using the polymerase chain reaction (PCR). The PCR product was cloned into pMD18-T vector, and subcloned into pET32a(+), generating the recombinant plasmid pET32a/DPV-gE. SDS-PAGE analysis showed that the fusion pET32a/DPV-gE protein was highly expressed after induction by 0.2 mM IPTG at 30°C for 4.5 h in Rosseta host cells. Over expressed 6×His-gE fusion protein was purified by nickel affinity chromatography, and used to immunize the rabbits for the preparation of polyclonal antibody. The result of the intracellular localization revealed that the gE protein was appeared to be in the cytoplasm region. The real time PCR, RT-PCR analysis and Western blotting revealed that the gE gene was produced most abundantly during the late phase of replication in DPV-infected cells. Conclusions. In this work, the DPV gE protein was successfully expressed in a prokaryotic expression system, and we presented the basic properties of the DPV gE product for the first time. These properties of the gE protein provided a prerequisite for further functional analysis of this gene. © 2010 Chang et al; licensee BioMed Central Ltd.


Xiang J.,Sichuan Agricultural University | Ma G.,China Rural Technology Development Center | Zhang S.,Sichuan Agricultural University | Cheng A.,Sichuan Agricultural University | And 10 more authors.
Virology Journal | Year: 2010

Knowledge of the intracellular location of a protein can provide useful insights into its function. Bioinformatic studies have predicted that the DEV pUL38 mainly targets the cytoplasm and nucleus. In this study, we obtained anti-pUL38 polyclonal sera. These antibodies were functional in western blotting and immunofluorescence in DEV-infected duck embryo fibroblasts (DEFs). pUL38 was expressed as a 51-kDa protein from 8 h post-infection onward, initially showing a diffuse distribution throughout the cytoplasm, and later in the nucleus. Furthermore, pUL38 was found in purified virus. These results provide the first evidence of the kinetics of expression and intracellular localization of DEV pUL38. © 2010 Xiang et al; licensee BioMed Central Ltd.

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