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Structural prediction of porcine sialoadhesin V-set Ig-like domain sheds some light on its role in porcine

Jie HOU,Rui LI,Hongfang MA,Songlin QIAO,Gaiping ZHANG

《农业科学与工程前沿（英文）》 2016年 第3卷 第1期   页码 65-71 doi: 10.15302/J-FASE-2016086

摘要： Porcine reproductive and respiratory syndrome (PRRS) is characterized by reproductive failures in sows and respiratory diseases in pigs of all ages. PRRS virus (PRRSV) is its causative agent and has caused huge economic losses in the swine industry. Porcine sialoadhesin (pSn) is a putative receptor of PRRSV. Previous studies have shown that a pSn V-set Ig-like domain is significant in PRRSV infection. However, its structural details are not fully known, hindering our deep understanding of PRRSV infection. In this study, we successfully cloned, expressed and purified the pSn V-set Ig-like domain in S2 cells. Then we tried to crystallize the target protein and predicted its structure. This will establish the foundation for the further structural study of pSn, deepen our understanding of the invasion mechanism of PRRSV, and support the structural information for the development of clinical drugs and vaccines against PRRSV.

关键词： PRRSV     porcine sialoadhesin     V-set Ig-like domain     Drosophila S2 cell     crystallization    

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Effect of calcium on porcine ICSI embryos expressing EGFP is related to activation of ooplasmic DNase

Shuaishuai WU,Heng CHEN,Yingzheng WANG,Hui GAO,Shenming ZENG

《农业科学与工程前沿（英文）》 2015年 第2卷 第1期   页码 84-89 doi: 10.15302/J-FASE-2015046

摘要： Several reliable methods to produce transgenic animals use the male genome. After penetration into oocytes, sperm DNA undergoes dramatic conformational changes that might represent an opportunity for exogenous DNA to integrate into the zygote genome. A nuclease, DNase I, with Ca /Mg dependent activity and Zn inhibition, is one of the enzymes responsible for sperm DNA remodeling. To date, the effect of different calcium concentrations in manipulation media on porcine intracytoplasmic sperm injection has not been fully investigated. The present study was conducted to examine the effect of calcium in the surrounding media, and we found that the number of embryos expressing green fluorescent protein (EGFP) was increased in media containing Ca . However, the number did not change over Ca concentrations from 2 to 8 mmol·L ( >0.05). Moreover, free Ca concentrations in the media were found to affect the efficiency which is ICSI embryos expressing EGFP protein, which was related to the activation of ooplasmic DNase I. These findings reveal a mechanism and pathway involving EGFP expression in ICSI embryos.

关键词： ICSI     calcium     DNase I     GFP     porcine    

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Progress, problems and prospects of porcine pluripotent stem cells

Hanning WANG,Yangli PEI,Ning LI,Jianyong HAN

《农业科学与工程前沿（英文）》 2014年 第1卷 第1期   页码 6-15 doi: 10.15302/J-FASE-2014009

摘要： Pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced PSCs (iPSCs), can differentiate into cells of the three germ layers, suggesting that PSCs have great potential for basic developmental biology research and wide applications for clinical medicine. Genuine ESCs and iPSCs have been derived from mice and rats, but not from livestock such as the pig—an ideal animal model for studying human disease and regenerative medicine due to similarities with human physiologic processes. Efforts to derive porcine ESCs and iPSCs have not yielded high-quality PSCs that can produce chimeras with germline transmission. Thus, exploration of the unique porcine gene regulation network of preimplantation embryonic development may permit optimization of culture systems for raising porcine PSCs. Here we summarize the recent progress in porcine PSC generation as well as the problems encountered during this progress and we depict prospects for generating porcine naive PSCs.

关键词： induced pluripotent stem cells     embryonic stem cells     pig     reprogramming    

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Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine

Jianwen CHEN, Kaiyuan PAN, Zhen CHEN, Biao DING, Dandan SONG, Wenbin BAO, Yunhai ZHANG

《农业科学与工程前沿（英文）》 2019年 第6卷 第1期   页码 66-72 doi: 10.15302/J-FASE-2018229

摘要：

Porcine viral diarrhea is an acute and highly contagious enteric disease of pigs that causes huge economic losses worldwide. Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) are the main pathogens responsible for piglet viral diarrhea. However, currently there is no specific drug available for the effective treatment of viral diarrhea. Therefore, it is necessary to seek an effective method to diminish PEDV and TGEV infection rates. RNA interference has been applied successfully to inhibit the virus replication. It provides a potential strategy for breeding resistant pigs. In this study, four promoters and four short hairpin RNA (shRNA) vectors with LoxP sites at each end of the selectable marker genes were constructed to target PEDV and TGEV. These vectors were then transfected into porcine fetal fibroblasts, G418 resistant transfectants were confirmed by PCR and transgenic SCNT porcine blastocysts were obtained. These results have paved the way for future production of marker-free transgenic resistant to PEDV and TEGV pigs by SCNT.

关键词： piglet diarrhea     RNAi     PEDV     TGEV     transgenic resistance breeding    

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SALL4 maintains self-renewal of porcine pluripotent stem cells through downregulation of OTX2

Ning WANG, Sile WANG, Yaxian WANG, Yuanxing CAI, Fan YANG, Huayan WANG

《农业科学与工程前沿（英文）》 2019年 第6卷 第1期   页码 81-92 doi: 10.15302/J-FASE-2017180

摘要：

Sall4 as one of the spalt family members contains several alternative splicing variants, which are differentially expressed and has a key role in maintaining pluripotent stem cells. However, the molecular features and function of SALL4 have not been well elucidated in porcine induced pluripotent stem cells (piPSCs). In this study, we identified splice variants and found two splicing variants through analysis of the porcine transcriptome data derived from piPSCs. SALL4A was only detected in piPSCs but SALL4B was globally expressed in porcine tissues and piPSCs. The level of SALL4B was significantly reduced when piPSCs differentiation occurred, however, the expression of SALL4A was not affected, indicating that SALL4B may be essential for the maintenance of piPSCs self-renewal. Overexpression of SALL4A and SALL4B in PEF cells could significantly stimulated expression of endogenous pluripotent genes, when SALL4B significantly promoted OCT4 expression. Conversely, SALL4A significantly promoted KLF4 expression. Additionally, both SALL4A and SALL4B could repress promoter activity in a dose-dependent manner. Conversely, OTX2 also negatively regulated SALL4 expression. These observations indicate that a negative feedback regulatory mechanism may exist between SALL4 and OTX2, which is useful for the maintenance of the self-renewal of piPSCs.

关键词： OTX2     pluripotency     pig     SALL4     transcription regulation    

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Characterization of polyclonal antibodies against nonstructural protein 9 from the porcine reproductive

Mengmeng ZHAO,Juanjuan QIAN,Jiexiong XIE,Tiantian CUI,Songling FENG,Guoqiang WANG,Ruining WANG,Guihong ZHANG

《农业科学与工程前沿（英文）》 2016年 第3卷 第2期   页码 153-160 doi: 10.15302/J-FASE-2016097

摘要： Porcine reproductive and respiratory syndrome (PRRS) is considered to be one of the most important infectious diseases impacting the swine industry and is characterized by reproductive failure in late term gestation in sows and respiratory disease in pigs of all ages. The nonstructural protein 9 gene, , encoding the RNA-dependent RNA polymerase, is generally regarded as fairly conserved when compared to other viral proteins. Antibodies against 9 will be of great importance for the diagnosis and treatment of the causal agent, PRRS virus. A study was undertaken to generate polyclonal antibodies against the immunodominant 9. For this purpose, the 9 was expressed in and subsequently used as an antigen to immunize New Zealand rabbits. Antiserum was identified via an indirect ELISA, and then verified based on the ability to react with both naturally and artificially expressed 9. Results of virus neutralization test showed that this antiserum could not neutralize the PRRSV. Nevertheless, this antiserum as a diagnostic core reagent should prove invaluable for further investigations into the mechanism of PRRS pathogenesis.

关键词： PRRSV     Nsp9     expression     antibody     neutralize    

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Transcriptomic basis of neutrophil ratio variation induced by poly I:C stimulation in porcine peripheral

Haiyan WANG, Qiaoxia ZHANG, Lilin YIN, Xiangdong LIU, Shuhong ZHAO, Mengjin ZHU, Changchun LI

《农业科学与工程前沿（英文）》 2017年 第4卷 第3期   页码 342-352 doi: 10.15302/J-FASE-2017162

摘要： Neutrophils are vital components of defense mechanisms against invading pathogens and are closely linked with the individual antiviral capacity of pigs and other mammals. Neutrophilia is a well-known clinical characteristic of viral and bacterial infections. Using Affymetrix porcine genome microarrays, we investigated the gene expression profiles associated with neutrophil variation in porcine peripheral blood before and after polyriboinosinic-polyribocytidylic acid stimulation. Transcriptomic analysis showed 796 differentially expressed genes (DEGs) in extreme response (ER) pigs and 192 DEGs in moderate response (MR) pigs. Most DEGs were related to immune responses, included , , , , , , , and . Gene ontology analysis indicated that the DEGs of both ER and MR pigs were involved in common biological processes, such as cell proliferation, growth regulation, immune response, inflammatory response and cell activation. The ER and MR groups also showed differences in DEGs involved in biological processes. DEGs involved in cell division and cell cycle were specifically found in the ER pigs, whereas DEGs involved in cell migration were specifically found in the MR pigs. The study provides a basic understanding of the molecular basis for the antiviral capacity of pigs and other mammals.

关键词： neutrophil     peripheral blood     pig     poly I:C     transcriptome    

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A brief review of microRNA and its role in PRRSV infection and replication

Xuekun GUO,Wenhai FENG

《农业科学与工程前沿（英文）》 2014年 第1卷 第2期   页码 114-120 doi: 10.15302/J-FASE-2014022

摘要： Porcine reproductive and respiratory syndrome virus (PRRSV), a single-stranded RNA virus, mainly infects cells of monocyte/macrophage lineage. Recently, host microRNAs were shown to be capable of modulating PRRSV infection and replication by multiple ways such as targeting viral genomic RNA, targeting viral receptor and inducing antiviral response. MicroRNAs are small RNAs and have emerged as important regulators of virus-host cell interactions. In this review, we discuss the identified functions of host microRNAs in relation to PRRSV infection and propose that cellular microRNAs may have a substantial effect on cell or tissue tropism of PRRSV.

关键词： porcine reproductive and respiratory syndrome virus (PRRSV)     microRNA     antiviral     viral tropism    

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Porcine pluripotent stem cells: progress, challenges and prospects

Jianyong HAN, Yi-Liang MIAO, Jinlian HUA, Yan LI, Xue ZHANG, Jilong ZHOU, Na LI, Ying ZHANG, Jinying ZHANG, Zhonghua LIU

《农业科学与工程前沿（英文）》 2019年 第6卷 第1期   页码 8-27 doi: 10.15302/J-FASE-2018233

摘要：

Pluripotent stem cells (PSCs) are characterized by their capacity for high self-renewal and multiple differentiation potential and include embryonic stem cells, embryonic germ cells and induced PSCs. PSCs provide a very suitable model for the studies of human diseases, drugs screening, regenerative medicine and developmental biology research. Pigs are considered as an ideal model for preclinical development of human xenotransplantation, therapeutic approaches and regenerative medicine because of their size and physiological similarity to humans. However, lack of knowledge about the derivation, characterization and pluripotency mechanisms of porcine PSCs hinders progress in these biotechnologies. In this review, we discuss the latest progress on porcine PSCs generation, evaluation criteria for pluripotency, the scientific and technical questions arising from these studies. We also introduce our perspectives on porcine PSC research, in the hope of providing new ideas for generating naive porcine PSCs and animal breeding.

关键词： embryonic germ cells     embryonic stem cells     induced pluripotent stem cells     pigs     pluripotent stem cells    

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标题 作者 时间 类型 操作

Structural prediction of porcine sialoadhesin V-set Ig-like domain sheds some light on its role in porcine

Jie HOU,Rui LI,Hongfang MA,Songlin QIAO,Gaiping ZHANG

期刊论文

Effect of calcium on porcine ICSI embryos expressing EGFP is related to activation of ooplasmic DNase

Shuaishuai WU,Heng CHEN,Yingzheng WANG,Hui GAO,Shenming ZENG

期刊论文

Progress, problems and prospects of porcine pluripotent stem cells

Hanning WANG,Yangli PEI,Ning LI,Jianyong HAN

期刊论文

Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine

Jianwen CHEN, Kaiyuan PAN, Zhen CHEN, Biao DING, Dandan SONG, Wenbin BAO, Yunhai ZHANG

期刊论文

SALL4 maintains self-renewal of porcine pluripotent stem cells through downregulation of OTX2

Ning WANG, Sile WANG, Yaxian WANG, Yuanxing CAI, Fan YANG, Huayan WANG

期刊论文

Characterization of polyclonal antibodies against nonstructural protein 9 from the porcine reproductive

Mengmeng ZHAO,Juanjuan QIAN,Jiexiong XIE,Tiantian CUI,Songling FENG,Guoqiang WANG,Ruining WANG,Guihong ZHANG

期刊论文

Transcriptomic basis of neutrophil ratio variation induced by poly I:C stimulation in porcine peripheral

Haiyan WANG, Qiaoxia ZHANG, Lilin YIN, Xiangdong LIU, Shuhong ZHAO, Mengjin ZHU, Changchun LI

期刊论文

A brief review of microRNA and its role in PRRSV infection and replication

Xuekun GUO,Wenhai FENG

期刊论文

Porcine pluripotent stem cells: progress, challenges and prospects

Jianyong HAN, Yi-Liang MIAO, Jinlian HUA, Yan LI, Xue ZHANG, Jilong ZHOU, Na LI, Ying ZHANG, Jinying ZHANG, Zhonghua LIU

期刊论文