Jianping Chen

Publications (35)85.27 Total impact

• Article: Experimental and bioinformatic evidence that RLBV P4 is a movement protein of the 30K superfamily.

  Chulang Yu, David Karlin, Yuwen Lu, Kathryn Wright, Jianping Chen, Stuart Macfarlane
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  ABSTRACT: Emaravirus is a recently described genus of negative-strand RNA plant viruses. Emaravirus P4 protein localizes to plasmodesmata, suggesting that it could be a viral movement protein (MP). In the current study, we showed that the P4 protein of Raspberry leaf blotch emaravirus (RLBV) rescues the cell-to-cell movement of a defective Potato virus X (PVX) that has a deletion mutation in the Triple Gene Block 1 (TGB1) movement-associated protein. This demonstrates that RLBV P4 is a functional MP. Sequence analyses revealed that P4 is a distant member of the 30K superfamily of MPs. All MPs of this family contain two highly conserved regions predicted to form β-strands, namely β1 and β2. We explored by alanine mutagenesis the role of two residues of P4 (Ile106 and Asp127) located in each of these strands. We also made the equivalent substitutions in the 29K MP of Tobacco rattle virus (TRV), another member of the 30K superfamily. All substitutions abolished the ability to complement PVX movement, except the I106A substitution in the β1 region of P4. This region has been shown to mediate membrane association of 30K MPs; our results show that it is possible to make non-conservative substitutions of a well conserved aliphatic residue within β1 without disrupting the membrane association or movement function of P4.
  Journal of General Virology 06/2013; · 3.36 Impact Factor
• Article: Transcription of ORFs on RNA2 and RNA4 of Rice stripe virus terminate at an AUCCGGAU sequence that is conserved in the genus Tenuivirus.

  Gentu Wu, Yuwen Lu, Hongying Zheng, Lin Lin, Fei Yan, Jianping Chen
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  ABSTRACT: Rice stripe virus, the type member of the genus Tenuivirus, has four genomic RNAs. RNAs 2-4 have an ambisense coding strategy and the noncoding intergenic regions (IRs) separating the two ORFs are thought to function in termination of transcription. Sequencing the 3' untranslated region of transcripts from RNA2 and RNA4 in virus-infected Oryza sativa (the natural host), Nicotiana benthamiana (an experimental host) and Laodelphax striatellus (the vector), showed that the sequences of p2 and pc2 transcripts on RNA2, and p4 and pc4 transcripts on RNA4 terminated with high frequency at a palindromic sequence AUCCGGAU that was located in a region predicted to form a hairpin secondary structure. The AUCCGGAU sequence is highly conserved in RNA2 and RNA4 of different RSV isolates and is also conserved among the corresponding genomic RNAs of other tenuiviruses. p3 transcripts from the three hosts all had the same dominant termination site, while pc3 transcripts from different hosts terminated at different sites. All pc1 3'UTR sequences ended at the 3' end of the viral complementary strand of RNA1 (data not shown), indicating that the pc1 transcript may be synthesized by runoff of viral polymerase, but had no characteristic termination sequence. This is the first experimental report determining the exact transcription termination sites of a plant ambisense virus, and has implications for understanding the transcription of RSV as well as other plant viruses with an ambisense coding strategy.
  Virus Research 04/2013; · 2.94 Impact Factor
• Article: Universal vectors for constructing artificial microRNAs in plants.

  Jie Zhou, Feibo Yu, Bin Chen, Xuming Wang, Yong Yang, Ye Cheng, Chengqi Yan, Jianping Chen
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  ABSTRACT: Universal amiRNA vectors (pUAs) for constructing plant amiRNAs in Arabidopsis and rice have been developed. By using type IIg restriction enzyme, BaeI, a single amiRNA construct can be produced using only one PCR and one ligation reaction. Thus, only one pair of primers is required for each amiRNA vector and these can be designed to be compatible with existing or newly developed methods. Because the BaeI recognition sequence is completely digested, there is no modification to the miRNA backbone, therefore avoids the risk of sequence changes that may affect downstream analysis. Based on these vectors, specific amiRNA constructs were created and verified. With optimized parameters, 38-45 % colonies for each amiRNA construct contain insertions with the expected orientation, and approximately 80 % of these colonies have the correct sequences.
  Biotechnology Letters 04/2013; · 1.68 Impact Factor
• Article: Enhanced transgene expression in rice following selection controlled by weak promoters.

  Jie Zhou, Yong Yang, Xuming Wang, Feibo Yu, Chulang Yu, Juan Chen, Ye Cheng, Chenqi Yan, Jianping Chen
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  ABSTRACT: BACKGROUND: Techniques that enable high levels of transgene expression in plants are attractive for the commercial production of plant-made recombinant pharmaceutical proteins or other gene transfer related strategies. The conventional way to increase the yield of desired transgenic products is to use strong promoters to control the expression of the transgene. Although many such promoters have been identified and characterized, the increase obtainable from a single promoter is ultimately limited to a certain extent. RESULTS: In this study, we report a method to magnify the effect of a single promoter by using a weak promoter-based selection system in transgenic rice. tCUP1, a fragment derived from the tobacco cryptic promoter (tCUP), was tested for its activity in rice by fusion to both a beta-glucuronidase (GUS) reporter and a hygromycin phosphotransferase (HPT) selectable marker. The tCUP1 promoter allowed the recovery of transformed rice plants and conferred tissue specific expression of the GUS reporter, but was much weaker than the CaMV 35S promoter in driving a selectable marker for growth of resistant calli. However, in the resistant calli and regenerated transgenic plants selected by the use of tCUP1, the constitutive expression of green fluorescent protein (GFP) was dramatically increased as a result of the additive effect of multiple T-DNA insertions. The correlation between attenuated selection by a weak promoter and elevation of copy number and foreign gene expression was confirmed by using another relatively weak promoter from nopaline synthase (Nos). CONCLUSIONS: The use of weak promoter derived selectable markers leads to a high T-DNA copy number and then greatly increases the expression of the foreign gene. The method described here provides an effective approach to robustly enhance the expression of heterogenous transgenes through copy number manipulation in rice.
  BMC Biotechnology 03/2013; 13(1):29. · 2.35 Impact Factor
• Article: Evaluation of High-Resolution Melting for Gene Mapping in Rice

  Jinshan Li, Xuming Wang, Ruixian Dong, Yong Yang, Jie Zhou, Chulang Yu, Ye Cheng, Chengqi Yan, Jianping Chen
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  ABSTRACT: In this study, high-resolution melting (HRM) analysis was evaluated for gene mapping in rice with sequence-tagged site (STS) and simple sequence repeat (SSR) markers. A total of 103 out of 353 normal STS and SSR markers revealed polymorphic melting curves among the parental genotypes, and 12 of these were successfully used to genotype the F2 mapping population for HRM analysis. Additional electrophoresis findings demonstrated that HRM genotyping matched with traditional electrophoresis results. To optimize the HRM-marker screening efficiency, different HRM reaction conditions were evaluated. A 5-μl touchdown-polymerase chain reaction (PCR) system provided no significant improvement in screening efficiency but in a 10-μl touchdown-PCR system, the marker screening efficiency increased by 75%. Twenty-one markers were obtained for mapping purposes under the optimized reaction conditions. This study indicates that HRM analysis can speed up the gene mapping progress in rice, while saving a lot of manpower. Keywords Oryza sativa –Gene mapping–High-resolution melting–Touchdown-PCR
  Plant Molecular Biology Reporter 04/2012; 29(4):979-985. · 2.45 Impact Factor
• Article: Genome organization and phylogenetic tree analysis of Garlic virus E, a new member of genus Allexivirus

  Jiong Chen, Jianping Chen
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  ABSTRACT: The complete sequence of an Allexivirus isolated from garlic plants in Yuhang City, Zhejiang Province, China had been determined. The single-strand, positive RNA genome was 8451 nucleotides in length excluding poly(A) tail. The genome organization of this virus was similar to that of the other Allexiviruses but only with 62.8%–64.8% nucleotide acid identities. The amino acid sequences of proteins encoded by ORF1-6 shared 67.6%–78.5%, 55.4%–66.2%, 56.7%–66.4%,40.3%–55.6%,66.3%–79.7%and 52.2%–68.8% identities with those of the others respectively. The homology range between it and the other Allexiviruses was similar to that between the other distinct species in this genus. A more comprehensive comparison using all available CP amino acid sequences showed that it shared only 63.9%–79.8% amino acids identical with the others. Therefore, it had been considered as a new member of the genus, named as garlic virus E (GarV-E). Phylogenetic analysis confirmed GarV-E as a distinct member and the correct names and classification of some members of genus Allexivirus were also discussed.
  Chinese Science Bulletin 04/2012; 47(1):33-37. · 1.32 Impact Factor
• Article: Sequence analysis of a soilborne wheat mosaic virus isolate from Italy shows that it is the same virus asEuropean wheat mosaic virus andSoilborne rye mosaic virus

  Jianping Yang, Jianping Chen, Jiong Chen, Ye Cheng, M. J. Adams
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  ABSTRACT: The complete sequence of the two RNAs of a furovirus isolate from durum wheat in Italy was determined. Sequence comparisons and phylogenetic analysis were done to compare the Italian virus withSoilborne wheat mosaic virus (SBWMV) from the USA and with furovirus sequences recently published asEuropean wheat mosaic virus (EWMV), from wheat in France, andSoilborne rye mosaic virus (SBRMV), from rye and wheat in Germany. Over the entire genome, the Italian isolate RNA1 and RNA2 had respectively 97.5% and 98.6% nucleotide identity with EWMV, 95.5% and 85.8% with SBRMV-G and 70.6% and 64.5% with SBWMV. The Italian isolate was therefore clearly distinct from SBWMV The European isolates all appear to belong to the same virus and the nameSoilborne cereal mosaic virus may resolve earlier ambiguities.
  Science in China Series C Life Sciences 04/2012; 44(2):216-224. · 1.61 Impact Factor
• Article: Two-step method for constructing Arabidopsis artificial microRNA vectors.

  Xuming Wang, Yong Yang, Jie Zhou, Chulang Yu, Ye Cheng, Chengqi Yan, Jianping Chen
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  ABSTRACT: Artificial microRNA (amiRNA) technology is used for gene silencing in Arabidopsis. We describe a method for constructing amiRNA vectors that requires only one PCR and one ligation reaction. Vectors produced by this method are the same as those from the method of Schwab et al. (Plant Cell 2006, 18:1121-1133). Transgenic plants created by this method can therefore be tested in the same way or compared with existing transgenic material without the risk of alteration to the amiRNA skeleton. With optimized parameters, 36-42 % colonies had the insertion in the expected orientation and 85-95 % of these had the correct sequence. Using this method, a transient gene knock-down analysis in Arabidopsis could be completed in 4-5 days.
  Biotechnology Letters 03/2012; 34(7):1343-9. · 1.68 Impact Factor
• Article: A simplified method for constructing artificial microRNAs based on the osa-MIR528 precursor.

  Fei Yan, Yuwen Lu, Gentu Wu, Jiejun Peng, Hongying Zheng, Lin Lin, Jianping Chen
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  ABSTRACT: Artificial miRNAs (amiRNAs) have been used successfully in various plants to silence endogenous genes or viral RNAs. The method of Schwab et al., widely used to construct amiRNAs, requires four PCRs. We describe a simplified method of constructing amiRNA based on the osa-MIR528 backbone using one PCR step by addition of prolonging sequence to the primers. The length of prolonging sequence needed in the osa-MIR528 precursor was determined. Using this method, we constructed amiRNA targeting the Nicotiana benthamiana UPF1 gene and showed that it functioned in silencing UPF1 expression in leaves when expressed transiently.
  Journal of biotechnology 03/2012; 160(3-4):146-50. · 2.88 Impact Factor
• Source

  Available from: Fei Yan

  Article: Identification of Novel Oryza sativa miRNAs in Deep Sequencing-Based Small RNA Libraries of Rice Infected with Rice Stripe Virus.

  Weixia Guo, Gentu Wu, Fei Yan, Yuwen Lu, Hongying Zheng, Lin Lin, Hairu Chen, Jianping Chen
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  ABSTRACT: MicroRNAs (miRNAs) play essential regulatory roles in the development of eukaryotes. Methods based on deep-sequencing have provided a powerful high-throughput strategy for identifying novel miRNAs and have previously been used to identify over 100 novel miRNAs from rice. Most of these reports are related to studies of rice development, tissue differentiation, or abiotic stress, but novel rice miRNAs related to viral infection have rarely been identified. In previous work, we constructed and pyrosequenced the small RNA (sRNA) libraries of rice infected with Rice stripe virus and described the character of the small interfering RNAs (siRNA) derived from the RSV RNA genome. We now report the identification of novel miRNAs from the abundant sRNAs (with a minimum of 100 sequencing reads) in the sRNA library of RSV-infected rice. 7 putative novel miRNAs (pn-miRNAs) whose precursor sequences have not previously been described were identified and could be detected by Northern blot or RT-PCR, and were recognized as novel miRNAs (n-miRNAs). Further analysis showed that 5 of the 7 n-miRNAs were up-expressed while the other 2 n-miRNAs were down-expressed in RSV-infected rice. In addition, 23 pn-miRNAs that were newly produced from 19 known miRNA precursors were also identified. This is first report of novel rice miRNAs produced from new precursors related to RSV infection.
  PLoS ONE 01/2012; 7(10):e46443. · 4.09 Impact Factor
• Source

  Available from: Fei Yan

  Article: The ability of PVX p25 to form RL structures in plant cells is necessary for its function in movement, but not for its suppression of RNA silencing.

  Fei Yan, Yuwen Lu, Lin Lin, Hongying Zheng, Jianping Chen
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  ABSTRACT: The p25 triple gene block protein of Potato virus X (PVX) is multifunctional, participating in viral movement and acting as a suppressor of RNA silencing. The cell-to-cell movement of PVX is known to depend on the suppression function of p25. GFP-fused p25 accumulates in rod-like (RL) structures with intense fluorescence in cells. By monitoring the location of fluorescence at different times, we have now shown that the RL structure is composed of filaments. P25 mutants without the conditional ability to recover movement function could not form RL structures while the mutants that had the ability did form the structure, suggesting that the ability of p25 to form RL structures is necessary for its function in cell-to-cell movement, but not for its suppressor function. Moreover, chemical inhibition of microfilaments in cells destroyed the formation of the complete RL structure. Additionally, TGBp2 and TGBp3 were recruited into the RL structure, suggesting a relationship between the TGBps in virus movement.
  PLoS ONE 01/2012; 7(8):e43242. · 4.09 Impact Factor
• Article: Garlic virus X 11-kDa protein granules move within the cytoplasm and traffic a host protein normally found in the nucleolus.

  Yuwen Lu, Fei Yan, Wei Guo, Hongying Zheng, Lin Lin, Jiejun Peng, Michael J Adams, Jianping Chen
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  ABSTRACT: The subcellular localization of the 11-kDa protein (p11) encoded by ORF3 of Garlic virus X (GarVX; genus Allexivirus, family Alphaflexiviridae) was examined by confocal microscopy. Granules with intense fluorescence were visible on the endoplasmic reticulum when p11 fused with green or red fluorescent protein (GFP or RFP) was expressed in epidermal cells of Nicotiana benthamiana. Moreover, the p11-RFP granules moved in the cytoplasm, along the cell periphery and through the cell membranes to adjacent cells. A 17-kDa protein (p17) of garlic interacting with p11 was identified by yeast two-hybridization and bimolecular fluorescence complementation assay. When p17 fused to GFP was expressed in epidermal cells of N. benthamiana, it localized to the nucleolus. However, in the presence of GarVX p11, the distribution of p17 changed to that of p11, but did not appear to affect the pattern of movement of p11.
  Molecular Plant Pathology 09/2011; 12(7):666-76. · 3.90 Impact Factor
• Article: Intron retention and 3'-UTR analysis of Arabidopsis Dicer-like 2 transcripts.

  Qiongji He, Jiejun Peng, Fei Yan, Lin Lin, Yuwen Lu, Hongying Zheng, Hairu Chen, Jianping Chen
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  ABSTRACT: Arabidopsis thaliana Dicer-like protein 2 (AtDCL2) plays an essential role in the RNA interference pathway. The function of AtDCL2 and other DCLs has been much studied but little has been done to characterize the DCLs transcripts before they are translated into proteins. Here, we investigated AtDCL2 transcripts and showed that all 21 introns of AtDCL2 except intron 9, 18, 20 and 21 could be retained although spliced sequences usually predominated. Intron 10 was more frequently retained and transient expression assays in Nicotiana benthamiana leaves showed that when AG/C at the 3' splicing site of the intron was changed to AG/G, the intron was more frequently spliced out. Conversely, a high retention of intron 18 was obtained if the AG/G at the 3' splicing site was changed to AG/C. These results suggest that the sequence at the 3' splicing site affects the efficiency of intron splicing. The 3'-UTRs of AtDCL2 had lengths between 54 and 154 nts, and the different 3'-UTRs differentially affected the transcriptional levels of fused GFP expressed transiently in N. benthamiana. Further comparisons and mutation experiments suggested that a putative SBF-1 binding site and an AU-rich element in the 3'-UTR both down-regulated expression of the upstream GFP fused to the 3'-UTR. Conversely, a second poly(A) consensus signal sequence in one 3'-UTR up-regulated gene expression. Our results provide insight into the character of AtDCL2 transcripts and demonstrate the potential complexity of factors that affect the frequency and patterns of alternative splicing.
  Molecular Biology Reports 06/2011; 39(3):3271-80. · 2.93 Impact Factor
• Article: Interaction between potyvirus P3 and ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) of host plants.

  Lin Lin, Zhaopeng Luo, Fei Yan, Yuwen Lu, Hongying Zheng, Jianping Chen
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  ABSTRACT: The P3 protein encoded by Shallot yellow stripe virus onion isolate (SYSV-O) interacted in the Yeast Two-hybrid (Y2H) system and in co-immunoprecipitation (Co-IP) assays with the large subunit of the ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) protein that is encoded by the rbcL gene of its onion host. Dissection analysis by Y2H showed that the main part of SYSV P3 (amino acids 1-390) and onion RbcL (amino acids 1-137) were responsible for the interaction. The P3 proteins encoded by Onion yellow dwarf virus (OYDV), Soybean mosaic virus Pinellia isolate (SMV-P), and Turnip mosaic virus (TuMV) also interacted with RbcL, suggesting that a P3/RbcL interaction might exist generally for potyviruses. An interaction between P3 of these potyviruses and the small subunit of RubisCO (RbcS) was also demonstrated. Moreover, the P3N-PIPO protein encoded by a newly identified open reading frame embedded within the P3 cistron also interacted with both RbcL and RbcS. It is possible that the potyvirus P3 protein affects the normal functions of RubisCO which thus contributes to symptom development.
  Virus Genes 03/2011; 43(1):90-2. · 1.85 Impact Factor
• Article: Silencing of NbXrn4 facilitates the systemic infection of Tobacco mosaic virus in Nicotiana benthamiana.

  Jiejun Peng, Jian Yang, Fei Yan, Yuwen Lu, Shanshan Jiang, Lin Lin, Hongying Zheng, Hairu Chen, Jianping Chen
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  ABSTRACT: The 5'-3' exoribonucleases (Xrns) play key roles in degradation and processing pathways of several classes of RNAs including mRNA, rRNA, miRNA and other small RNAs. Recent work revealed that the cytoplasmic Xrn (Xrn1p in yeast and Xrn4 in plants) affected the stability of the viral RNA of tombusviruses in yeast and plants, which indicates that the cytoplasmic Xrn might be involved in plant defense against virus by degrading viral RNA. Here, we demonstrated that silencing of Nicotiana benthamiana cytoplasmic Xrn4 facilitated both local and systemic infection of Tobacco mosaic virus (TMV) in N. benthamiana. The results support the suggestion that cytoplasmic Xrn4 participates in the viral defense system of plants.
  Virus Research 03/2011; 158(1-2):268-70. · 2.94 Impact Factor
• Article: Mapping the self-interacting domains of TuMV HC-Pro and the subcellular localization of the protein.

  Hongying Zheng, Fei Yan, Yuwen Lu, Liying Sun, Lin Lin, Li Cai, Mingsheng Hou, Jianping Chen
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  ABSTRACT: The helper component-proteinase (HC-Pro) of potyviruses is a multifunctional protein involved in aphid transmission, polyprotein processing, cell-to-cell and long-distance movement, genome amplification and symptom expression. The HC-Pros of several potyviruses interact with themselves but the key domains responsible for self-interaction are apparently not conserved. In our experiments, yeast two-hybrid assays and bimolecular fluorescence complementation showed that Turnip mosaic virus (TuMV) HC-Pro interacted with itself in yeast cells, plant cells and insect cells. It was also shown that the central and C-terminal regions of the HC-Pro participated in these self-interactions. Fluorescence microscopy showed that TuMV HC-Pro was present in the cytoplasm and formed aggregates along the ER.
  Virus Genes 10/2010; 42(1):110-6. · 1.85 Impact Factor
• Article: Characterization of siRNAs derived from rice stripe virus in infected rice plants by deep sequencing.

  Fei Yan, Hengmu Zhang, Michael J Adams, Jian Yang, Jiejun Peng, John F Antoniw, Yijun Zhou, Jianping Chen
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  ABSTRACT: RNA interference is a natural defense against viruses in plants. To date, the only viral siRNAs characterized have been those for positive-sense RNA viruses with one or two genome components. Here, we characterized siRNAs derived from rice stripe virus (RSV), a member of the genus Tenuivirus with four genomic RNAs and an ambisense coding strategy. Deep sequencing of small RNAs from infected rice leaves showed that siRNAs were derived almost equally from virion and complementary RNA strands and were mostly 20-22 nucleotides long. Most viral siRNAs were produced within the coding sequences and 5' termini of the RSV genome. RSV siRNAs had a higher G and lower C content than the viral genome but a strong A/U bias at the first nucleotide and a U bias at the final one, suggesting preferential targeting of such sequences by rice Dicer-like proteins.
  Archives of Virology 06/2010; 155(6):935-40. · 2.11 Impact Factor
• Article: A highly efficient method for construction of rice artificial MicroRNA vectors.

  Xuming Wang, Yong Yang, Chulang Yu, Jie Zhou, Ye Cheng, Chengqi Yan, Jianping Chen
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  ABSTRACT: Artificial microRNA (amiRNA) has become a powerful tool for gene silencing in plants. A new method for easy and rapid construction of rice artificial miRNA vector is described. The procedure involved modification of the pCAMBIA1300-UR vector by insertion of a 'vector modification fragment'. This was prepared from the precursor of Os-amiR528 by eliminating the central miRNA-containing region while simultaneously creating an AfeI restriction site. The fragment was then introduced to the destination vector to produce a multipurpose 'Highly Efficient gene Silencing Compatible vector' (HESC vector). AfeI was used to produce linearized HESC vectors, and a blunt end PCR product that included amiRNA sequence was cloned into this site by a single ligation reaction to create the completed amiRNA vector. Tests showed that the method was highly efficient, and greatly reduced the time needed for vector construction and resulted in a DNA sequence identical to that of the current method, making it particularly suitable for use in a systems biology approach to functional genomic research.
  Molecular Biotechnology 05/2010; 46(3):211-8. · 2.17 Impact Factor
• Article: Minor structural protein VP2 in rabbit hemorrhagic disease virus downregulates the expression of the viral capsid protein VP60.

  Liu Chen, Guangqing Liu, Zheng Ni, Bin Yu, Tao Yun, Yi Song, Jionggang Hua, Shuangmao Li, Jianping Chen
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  ABSTRACT: Rabbit hemorrhagic disease virus (RHDV) has two structural proteins: the major capsid protein VP60 and the minor capsid protein VP2. VP2 is speculated to play an important role in the virus life cycle. To investigate the effect of VP2 on VP60 expression, three types of experiment (baculovirus-insect cell system, mammalian-luciferase assay system and in vitro coupled transcription/translation system) were used to express VP60 alone or co-expressed with VP2. Both forms of VP60 were able to form virus-like particles in insect cells. Western blot analysis and dual-luciferase assays demonstrated that the presence of VP2 results in downregulation of the expression of VP60 in vivo. Real-time RT-PCR of mRNA levels showed that downregulation of VP60 occurs at the transcriptional level. The ability of the viral minor structural protein VP2 to regulate capsid protein levels may contribute to effective virus infection.
  Journal of General Virology 10/2009; 90(Pt 12):2952-5. · 3.36 Impact Factor
• Source

  Available from: Jiong Chen

  Article: Protein-protein interactions in two potyviruses using the yeast two-hybrid system.

  Lin Lin, Yuhong Shi, Zhaopeng Luo, Yuwen Lu, Hongying Zheng, Fei Yan, Jiong Chen, Jianping Chen, M J Adams, Yunfeng Wu
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  ABSTRACT: Interactions between all ten mature proteins of the potyviruses Soybean mosaic virus (Pinellia ternata isolate) and Shallot yellow stripe virus were investigated using yeast two-hybrid (Y2H) assays. Consistently strong self-interactions were found between the pairs of HC-Pro, VPg, NIa-Pro, NIb and CP in both viruses. Apart from the NIb, such interactions have been previously reported for some other potyviruses. The 6K1/NIa-Pro combination gave a consistently moderate to strong interaction in both directions for both viruses. This interaction occurred even when the 6K1 of SMV-P was truncated to eliminate the C-terminal motif that acts as a recognition site for cleavage by the NIa-Pro. Many other interactions occurred only in one direction or only for one of the two viruses. When taken together with other published reports, the data suggest that interactions detected by Y2H should be regarded as only preliminary indications.
  Virus Research 02/2009; 142(1-2):36-40. · 2.94 Impact Factor