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Synthesis of Reassortant Infectious Bursal Disease Virus in Chickens Injected Directly with Infectious Clones from Different Virus Strains

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Acta Biochimica et Biophysica Sinica 2005年第3期37卷 192-198页

作者： long li yao-wei huang lian-sheng wang wang-jun wan lian yu zhejiang provincial key laboratory of preventive veterinary medicine,institute of preventive veterinary medicine.zhejiang university.hangzhou 310029,china zhejiang provincial key laboratory of preventive veterinary medicine Institute of Preventive Veterinary Medicine Zhejiang UniversityHangzhou 310029 China

The infectious bursal disease virus（IBDV）,a member of the Birnaviridae family,containinga bisegmented double-stranded RNA genome,encodes four structural viral proteins,VP1,VP2,VP3,andVP4,as well as a non-structural protein,VP5,In the present paper,the segment A from two IBDV strains,field isolate ZJ2000 and attenuated strain HZ2,were inserted into one NaeI site by site-directed silent mu-tagenesis and subcloned into the eukaryotic expression plasmid pCI under the control of the human cy-tomegalovirus（hCMV）immediate early enhancer and promoter to construct the recombinant plasmids pCI-AKZJ2000and pCI-AKHZ2,*** of the two recombinants was combined with another recombinant pCIplasroid containing the marked segment B of strain HZ2（pCI-mB）,and injected intramuscularly into non-immunized *** chimeric IBDV strains were recovered from the *** out of eightchickens in each of two groups showed the bursal histopathological *** reassortant virus derivedfrom pCI-AKZJ2000/pCI-mB can infect chicken embryos and shows relatively low *** havedeveloped a novel virus reverse genetic approach for the study of *** results also form the basis forinvestigating the role of VP1 in viral replication and pathogenecity.

关键词： infectious bursal disease infectious bursal disease virus reverse genetics reassortant virus animal model

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Nitric Oxide Inducing Function and Intracellular Movement of Chicken Interleukin-18 in Cultured Cells

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Acta Biochimica et Biophysica Sinica 2005年第10期37卷 688-693页

作者： Jian XU~(1,3) Tong-Le DENG~2 long li~1 Zhen-Qiang YOU~1 wang-jun wan~2 lian yu~(1*) 1 zhejiang provincial key laboratory of preventive veterinary medicine.institute of preventive veterinary medicine.zhejiang university.hangzhou 310029.china 2 College of Biomedical Engineering and Instrument Science,zhejiang university.hangzhou 310029,china 3 Medical College of Shihezi university,Xinjiang 832002,china zhejiang provincial key laboratory of preventive veterinary medicine Institute of Preventive Veterinary Medicine Zhejiang UniversityHangzhou 310029 China Medical College of Shihezi universityXinjiang 832002 China College of Biomedical Engineering and Instrument Science Zhejiang UniversityHangzhou 310029 China

To evaluate the characteristics of chicken interleukin-18 (ChIL-18) in different forms in vitro,the ChIL-18 full-length gene (ChIL-18-F) and the ChIL-18 presumed mature protein gene (ChIL-18-M) werecloned and inserted into the eukaryotic expression vector pCI,to construct recombinant pCI-ChIL-18-Fand *** recombinant plasmids were then transferred into chicken splenic lymphocytes(CSLs).Western blot showed that ChIL-18-F,with a molecular weight of 23.0 kDa,was produced in CSLstransfected by pCI-ChlL-18-F;ChIL-18-M,with a molecular weight of 19.5 kDa,was produced in CSLstransfected by *** nitric oxide (NO) level in the transfected CSLs and the culture mediumat different time points was further examined under confocal microscopy using *** results showed that both pCI-ChIL-18-F and pCI-ChIL-18-M groups showed significantincrease in intracellular and extracellular NO production compared with pCI transfected control *** suggest that both ChIL-18-F and ChIL-18-M could stimulate NO secretion in *** characterizethe intracellular distribution of ChIL-18,ChIL-18-F and ChIL-18-M were each fused to the enhanced greenfluorescent protein gene,and expressed in Vero *** results showed that the ChIL-18-F tended to themembranous region in Vero cells,while ChIL-18-M did *** indicates that the N-terminal 27 amino acidpeptide helped ChIL-18 target to Veto cell membranes.

关键词： chicken interleukin-18 N-terminal peptide nitric oxide splenic lymphocyte intracellular movement

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