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| 1 | Construction of infectious cDNA clones of PRRSV:Separation of coding regions for nonstructural and structural proteins显示文摘Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), the causative agent of the ongoing 'porcine high fever syndrome' in China, is capable of genetic and antigenic mutations at high fre- quency. How to design vaccine rationally to keep up with the ever-changing prevalent PRRSV variant is of great interest. We developed an infectious cDNA clone of an attenuated strain of Type II PRRSV, and further manipulated the infectious cDNA clone by inserting polylinker between ORF1 and ORF2, en- coding for nonstructural- or structural-protein, respectively. The cDNA was generated from the cell-attenuated virus strain, APRRS, via RT-PCR, and followed by nucleotide sequencing and molecular cloning. The full-length of the APRRS genomic RNA was determined as 15521 nucleotides in length excluding poly(A) tail, which has a 99.7% nucleotide identity with that of PRRSV Nsp strain, also a vac- cine strain. Based on the nucleotide sequencing results, the full-length cDNA clone was assembled in pBlueScript vector, under the control of T7 promoter at the immediate 5′ terminus of genome. To dis- cern the rescued viruses from that of parental virus, a Mlu I restriction site was engineered into ORF5 coding region. Upon transfection of the in vitro transcripts of both the original and Mlu I-tagged cDNAs into MA-104 cells, typical PRRSV cytopathic effects were observed. The rescued viruses from the full-length cDNA clones displayed the same virological and molecular properties. Subsequently, PCR-based mutagenesis was conducted to separate the coding regions between PRRSV nonstructural genes, ORF1, and structural proteins, ORF2-ORF7. The synthetic RNA of such mutant clone, pCSA, was infectious and the rescued virus shared similar properties with that of the parental virus. This study provided a valuable tool for development of chimeric PRRSV as vaccine candidate offering cross-protection to various genetically diversified PRRSV strains, and a platform for further develop- ment of PRRSV as a gene expression vector for recombinant vaccines against other significant swine diseases. | YUAN ShiShan & WEI ZuZhang Department of Animal Infectious Diseases, Key Laboratory of Animal Parasitology Chinese Ministry of Agriculture, Shanghai Vet- erinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200232, China | 2008 | Science China(Life Sciences)2008，51，3： | 19 |
| 2 | Effects of long-term fertilization with different substitution ratios of organic fertilizer on paddy soil显示文摘In recent years,the abuse of chemical fertilizers has caused numerous environmental problems,such as soil acidification and compaction.Replacing chemical fertilizers with organic fertilizers can effectively alleviate these problems.However,the effects of alternative organic fertilizers remain unclear.To explore the effects of organic fertilizer substitution on rice yield and paddy soil physicochemical properties and bacterial community structure,we conducted a 5-year experiment using different proportions of organic fertilizer substitution in a double-cropping rice field in Jiangxi,China.Our results showed that replacing chemical fertilizers with organic fertilizers can reduce soil acidification,increase soil organic matter content,nutrient contents,and enzyme activities,improve soil physicochemical properties and microbial community,and enhance soil metabolism.Appropriate organic substitution also had positive effects on rice production.These findings enhance our understanding of the effects of different alternative organic fertilization methods and have important theoretical significance for the promotion of the use of organic fertilizers in the future. | Weifeng SONG Aiping SHU Jiai LIU Wenchong SHI Mingcong LI Wenxue ZHANG Zuzhang LI Guangrong LIU Fusheng YUAN Shuxin ZHANG Zengbing LIU Zheng GAO | 2022 | Pedosphere2022，32，4： | 7 |
| 3 | Construction and characterization of a full-length infectious clone of Getah virus in vivo显示文摘Getah virus(GETV)is a mosquito-borne virus of the genus Alphavirus in the family Togaviridae and,in recent years,it has caused several outbreaks in animals.The molecular basis for GETV pathogenicity is not well understood.Therefore,a reverse genetic system of GETV is needed to produce genetically modified viruses for the study of the viral replication and its pathogenic mechanism.Here,we generated a CMV-driven infectious cDNA clone based on a previously isolated GETV strain,GX201808(pGETV-GX).Transfection of pGETV-GX into BHK-21 cells resulted in the recovery of a recombinant virus(rGETV-GX)which showed similar growth characteristics to its parental virus.Then three-day-old mice were experimentally infected with either the parental or recombinant virus.The recombinant virus showed milder pathogenicity than the parental virus in the mice.Based on the established CMV-driven cDNA clone,subgenomic promoter and two restriction enzyme sites(BamHI and EcoRI)were introduced into the region between E1 protein and 3’UTR.Then the green fluorescent protein(GFP),red fluorescent protein(RFP)and improved light-oxygen-voltage(iLOV)genes were inserted into the restriction enzyme sites.Transfection of the constructs carrying the reporter genes into BHK-21 cells proved the rescue of the recombinant reporter viruses.Taken together,the establishment of a reverse genetic system for GETV provides a valuable tool for the study of the virus life cycle,and to aid the development of genetically engineered GETVs as vectors for foreign gene expression. | Tongwei Ren Xiangling Min Qingrong Mo Yuxu Wang Hao Wang Ying Chen Kang Ouyang Weijian Huang Zuzhang Wei | 2022 | Virologica Sinica2022，37，3： | 2 |
| 4 | High prevalence of a novel porcine bocavirus in weanling piglets with respiratory tract symptoms in China 显示文摘 | Zhai Shaolun Yue Cheng Wei Zuzhang | 2010 | Arch Virol2010，155，： | 1 |
| 5 | Long-term effects of organic amendments on the rice yields for double rice cropping systems in subtropical China显示文摘 | Lidong Bi Bin Zhang Guangrong Liu Zuzhang Li Yiren Liu Chuan Ye Xichu Yu Tao Lai Jiguang Zhang Jianmin Yin Yin Liang | 2008 | Agriculture Ecosystems and Environment2008，，4： | 1 |
| 6 | N-Linked Glycosylation of GP5 of Porcine Reproductive and Respiratory Syndrome Virus Is Critically Important for Virus Replication In Vivo显示文摘 | Zuzhang Wei Tao Lin Lichang Sun Yanhua Li Xiaoming Wang Fei Gao Runxia Liu Chunyan Chen Guangzhi Tong Shishan Yuan | 2012 | Journal of Virology2012，，18： | 1 |
| 7 | Immunization of pigs with a type 2 modified live PRRSV vaccine prevents the development of a deadly long lasting hyperpyrexia in a challenge study with highly pathogenic PRRSV JX143 显示文摘 | Zuzhang W Jianwu Zh Jinshan Zh | 2013 | Vaccine2013，，31： | 1 |
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