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Hu, Jianjun,Zhang, Wanqi,Chauhan, Surinder Singh,Shi, Changqing,Song, Yumeng,Zhao, Yubing,Wang, Zhehong,Cheng, Long,Zhang, Yingyu The Korean Society of Veterinary Science 2020 Journal of Veterinary Science Vol.21 No.2
Background: Bovine papilloma is a neoplastic disease caused by bovine papillomaviruses (BPVs), which were recently divided into 5 genera and at least 24 genotypes. Objectives: The complete genome sequence of BPV type 15 (BPV Aks-02), a novel putative BPV type from skin samples from infected cows in Southern Xinjiang China, was determined by collecting warty lesions, followed by DNA extraction and amplicon sequencing. Methods: DNA was analyzed initially by polymerase chain reaction (PCR) using the degenerate primers FAP59 and FAP64. The complete genome sequences of the BPV Aks-02 were amplified by PCR using the amplification primers and sequencing primers. Sequence analysis and phylogenetic analysis were performed using bio-informatic software. Results: The nucleotide sequence of the L1 open reading frame (ORF) of BPV Aks-02 was 75% identity to the L1 ORF of BPV-9 reference strain from GenBank. The complete genome consisted of 7,189 base pairs (G + C content of 42.50%) that encoded 5 early (E8, E7, E1, E2, and E4) and 2 late (L1 and L2) genes. The E7 protein contained a consensus CX<sub>2</sub>CX<sub>29</sub>CX<sub>2</sub>C zinc-binding domain and a LxCxE motif. Among the different members of this group, the percentages of the complete genome and ORFs (including 5 early and 2 late ORFs) sequence identity of BPV Aks-02 were closer to the genus Xipapillomavirus 1 of the Xipapillomavirus genus. Phylogenetic analysis and sequence similarities based on the L1 ORF of BPV Aks-02 revealed the same cluster. Conclusions: The results suggest that BPV type (BPV Aks-02) clustered with members of the Xipapillomavirus genus as BPV 15 and were closely related to Xipapillomavirus 1.
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Protective Immune Response of Bacterially-Derived Recombinant FaeG in Piglets
Huang Yahong,Wanqi Liang,Aihu Pan,Zhiai Zhou,Qiang Wang,Cheng Huang,Jianxiu Chen,Dabing Zhang 한국미생물학회 2006 The journal of microbiology Vol.44 No.5
FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli(ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, C83907 (K88ad, CT+, ST+). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.
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Jianjun Hu,Wanqi Zhang,Surinder Singh Chauhan,Changqing Shi,Yumeng Song,Yubing Zhao,Zhehong Wang,Long Cheng,Yingyu Zhang 대한수의학회 2020 Journal of Veterinary Science Vol.21 No.6
Background: Bovine papilloma is a neoplastic disease caused by bovine papillomaviruses (BPVs), which were recently divided into 5 genera and at least 24 genotypes. Objectives: The complete genome sequence of BPV type 15 (BPV Aks-02), a novel putative BPV type from skin samples from infected cows in Southern Xinjiang China, was determined by collecting warty lesions, followed by DNA extraction and amplicon sequencing. Methods: DNA was analyzed initially by polymerase chain reaction (PCR) using the degenerate primers FAP59 and FAP64. The complete genome sequences of the BPV Aks-02 were amplified by PCR using the amplification primers and sequencing primers. Sequence analysis and phylogenetic analysis were performed using bio-informatic software. Results: The nucleotide sequence of the L1 open reading frame (ORF) of BPV Aks-02 was 75% identity to the L1 ORF of BPV-9 reference strain from GenBank. The complete genome consisted of 7,189 base pairs (G + C content of 42.50%) that encoded 5 early (E8, E7, E1, E2, and E4) and 2 late (L1 and L2) genes. The E7 protein contained a consensus CX2CX29CX2C zinc-binding domain and a LxCxE motif. Among the different members of this group, the percentages of the complete genome and ORFs (including 5 early and 2 late ORFs) sequence identity of BPV Aks-02 were closer to the genus Xipapillomavirus 1 of the Xipapillomavirus genus. Phylogenetic analysis and sequence similarities based on the L1 ORF of BPV Aks-02 revealed the same cluster. Conclusions: The results suggest that BPV type (BPV Aks-02) clustered with members of the Xipapillomavirus genus as BPV 15 and were closely related to Xipapillomavirus 1.
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Protective Immune Response of Bacterially-Derived Recombinant FaeG in Piglets
Yahong, Huang,Liang, Wanqi,Pan, Aihu,Zhou, Zhiai,Wang, Qiang,Huang, Cheng,Chen, Jianxiu,Zhang, Dabing The Microbiological Society of Korea 2006 The journal of microbiology Vol.44 No.5
FaeG is the key factor in the infection process of K88ad enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin. In an attempt to determine the possibility of expressing recombinant FaeG with immunogenicity for a new safe and high-production vaccine in E. coli, we constructed the recombinant strain, BL21 (DE3+K88), which harbors an expression vector with a DNA fragment of faeG, without a signal peptide. Results of 15% SDS-polyacrylamide slab gel analysis showed that FaeG can be stably over-expressed in BL21 (DE3+K88) as inclusion bodies without FaeE. Immunoglobulin G (IgG) and M (IgM) responses in pregnant pigs, with boost injections of the purified recombinant FaeG, were detected 4 weeks later in the sera and colostrum. An in vitro villius-adhesion assay verified that the elicited antibodies in the sera of vaccinated pigs were capable of preventing the adhesion of K88ad ETEC to porcine intestinal receptors. The protective effect on the mortality rates of suckling piglets born to vaccinated mothers was also observed one week after oral challenge with the virulent ETEC strain, $C_{83907}$ (K88ad, $CT^+,\;ST^+$). The results of this study proved that the adhesin of proteinaceous bacterial fimbriae or pili could be overexpressed in engineered E. coli strains, with protective immune responses to the pathogen.
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Lu Zhu,Jianxin Shi,Guochao Zhao,Dabing Zhang,Wanqi Liang 한국식물학회 2013 Journal of Plant Biology Vol.56 No.1
The tapetum of the anther locule encloses the malereproductive cells and plays a supportive role for normalpollen development. However, the underlying mechanismremains less understood. Previously, we identified a completerecessive male sterile mutant, post-meiotic deficient anther1(pda1), with abnormal postmeiotic tapetal development. Inthis study we comprehensively characterized pda1. Chemicalanalysis uncovered that pda1 anther had significant lowerlevels of cutin monomers and cuticular waxes. PDA1 geneencodes an ATP-binding cassette (ABC) half-transporter,namely OsABCG15, which is conserved from algae tohigher plants. In situ RNA hybridization assay showed thatPDA1 is strongly expressed in tapetal cells, and weakly inmicrospores during the anther development. Additionally,the expression of two pollen exine biosynthetic genesCYP704B2 and CYP703A3 was dramatically reduced inpda1 mutant anthers. Altogether, these observations suggestthat the tapetum-expressed ABC transporter PDA1 plays acrucial role in secreting lipidic precursors from the tapetumto developing microspores and the anther epidermis.
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Xiaoling Li,Ligang Xing,Yujun Zhang,Peng Xie,Wanqi Zhu,Xiangjiao Meng,Yinxia Wang,Lingling Kong,Hanxi Zhao,Jinming Yu 한국식품영양과학회 2020 Journal of medicinal food Vol.23 No.1
Acute radiation-induced esophagitis (ARIE) is among the most serious form of toxicities associated with definitive radiotherapy or chemoradiotherapy used for treatment of patients with esophageal cancer. Our preliminary phase I and II trials of lung cancer patients who received radiotherapy indicated epigallocatechin-3-gallate (EGCG) as a promising therapeutic option against ARIE. Therefore, we conducted a prospective phase II study to validate the efficacy and safety of EGCG in the treatment of ARIE. The patients who received chemoradiotherapy or definitive radiotherapy for treatment of esophageal cancer in the Shandong Cancer Hospital and Institute in China were enrolled for the present study. EGCG (440 μM) was administered with first onset of ARIE and then at weeks after final radiotherapy. The patients were monitored every week for dysphagia, Radiation Therapy Oncology Group (RTOG) score, and esophagitis-related pain. Moreover, tumor response and the effect on survival following the treatment were also evaluated. Comparison of the RTOG score in the first, second, third, fourth, fifth, and even sixth week after EGCG prescription and the first and second week after radiotherapy with baseline indicates a significant reduction. The tumor response rate was 86.3%. The overall survival rate in 1, 2, and 3 years was found to be 74.5%, 58%, and 40.5%. Oral administration of EGCG solution seems to be feasible for treating ARIE in patients with esophageal cancer who receive radiation therapy. EGCG might be an ARIE-reliever without compromising the efficacy of radiation therapy. A randomized study with a control group is needed for further evaluation.
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Tissue-Specific CRISPR/Cas9 System of Cotton Pollen with GhPLIMP2b and GhMYB24 Promoters
Lei Jianfeng,Dai Peihong,Li Jiyang,Yang Mi,Li Xiuqin,Zhang Wanqi,Zhou Guantong,WangzhenGuo,Liu Xiaodong 한국식물학회 2021 Journal of Plant Biology Vol.64 No.1
CRISPR/Cas9 technology is a powerful tool for improving crop genetic traits. However, CRISPR/Cas9 system for efficient expression of cotton germ cells has still not been established. In this study, we developed a tissue-specific vectors to drive Cas9 expression with GhPLIMP2b and GhMYB24 promoters and established the effective method to transform cotton pollen by Agrobacterium vacuum infiltration. GhPLIMP2b and GhMYB24 promoters of cotton pollen were cloned into Cas9 expression vectors. The sgRNAs targetting to CLA1, ERA1 and GGB (drought-resistant negative regulation genes) were designed and constructed into GhPLIMP2b and GhMYB24 promoter vectors. Cotton pollens were tranformed by Agrobacterium vacuum infiltration with GhPLIMP2b and GhMYB24 promoter vectors. The results of clone sequencing shown that mutation types of the sequence were mainly base substitutions wth the frequency from 3.29 to 6.45%. Eleven potential off-target sites were chosen and two sites were observed. Our results indicated that GhPLIM2bP::Cas9 and GhMYB24P::Cas9 editing vectors achieved targeted edition of endogenous genes in cotton pollen, but there were a few off-target effects. This study provides an effective gene-editing system for the rapid acquisition of cotton mutants using pollen as a transgenic receptor.
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Yi, Jakyung,Moon, Sunok,Lee, Yang-Seok,Zhu, Lu,Liang, Wanqi,Zhang, Dabing,Jung, Ki-Hong,An, Gynheung American Society of Plant Biologists 2016 Plant Physiology Vol.170 No.3
<P>After meiosis, tapetal cells in the innermost anther wall layer undergo program cell death (PCD)-triggered degradation. This step is essential for microspore development and pollen wall maturation. We identified a key gene, Defective Tapetum Cell Death 1 (DTC1), that controls this degeneration by modulating the dynamics of reactive oxygen species (ROS) during rice male reproduction. Mutants defective in DTC1 exhibit phenotypes of an enlarged tapetum and middle layer with delayed degeneration, causing male sterility. The gene is preferentially expressed in the tapetal cells during early anther development. In dtc1 anthers, expression of genes encoding secretory proteases or lipid transporters is significantly reduced, while transcripts of PCD regulatory genes, e.g. UDT1, TDR1, and EAT1/DTD, are not altered. Moreover, levels of DTC1 transcripts are diminished in udt1, tdr, and eat1 anthers. These results suggest that DTC1 functions downstream of those transcription factor genes and upstream of the genes encoding secretory proteins. DTC1 protein interacts with OsMT2b, a ROS scavenger. Whereas wild-type plants accumulate large amounts of ROS in their anthers at Stage 9 of development, those levels remain low during all stages of development in dtc1 anthers. These findings indicate that DTC1 is a key regulator for tapetum PCD by inhibiting ROS-scavenging activity.</P>
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