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cloning pig

Anne Bugge, Peter Gildsig Jansen, Leonardo De Maria, Samra Joke Sanni, Trine Ryberg Clausen
The gastrointestinal hormone cholecystokinin (CCK) regulates digestive processes and satiety in addition to centrally mediated effects on nociception and anxiety. CCK signals through two seven-trans-membrane receptors named the CCK-1 receptor and the CCK-2 receptor. The expression pattern and biological effects mediated by the CCK-1 and CCK-2 receptors are highly divergent. The pig is a widely used preclinical animal model in medical research, but up until recently, the porcine CCK-2 receptor was described as a pseudogene in the publicly available genomic sequence databases...
June 16, 2018: European Journal of Pharmacology
Fengjuan Liu, Jinji Liu, Zaimei Yuan, Yubo Qing, Honghui Li, Kaixiang Xu, Wanyun Zhu, Heng Zhao, Baoyu Jia, Weirong Pan, Jianxiong Guo, Xuezeng Zhang, Wenmin Cheng, Wei Wang, Hong-Ye Zhao, Hong-Jiang Wei
Pig-to-human organ transplantation has drawn attention in recent years due to the potential use of pigs as an alternative source of human donor organs. While GGTA1 knockout (GTKO) can protect xenografts from hyperacute rejection, complement-dependent cytotoxicity might still contribute to this type of rejection. To prolong the xenograft survival, we utilized a T2A-mediated pCMV-hCD55-T2A-hCD59-Neo vector and transfected the plasmid into GTKO Diannan miniature pig fetal fibroblasts. After G418 selection combined with single-cell cloning culture, four colonies were obtained, and three of these were successfully transfected with the hCD55 and hCD59...
June 18, 2018: Molecular Biotechnology
Nathaniel Mon Père, Tom Lenaerts, Jorge M Pacheco, David Dingli
Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal blood disorder characterized by hemolysis and a high risk of thrombosis, that is due to a deficiency in several cell surface proteins that prevent complement activation. Its origin has been traced to a somatic mutation in the PIG-A gene within hematopoietic stem cells (HSC). However, to date the question of how this mutant clone expands in size to contribute significantly to hematopoiesis remains under debate. One hypothesis posits the existence of a selective advantage of PIG-A mutated cells due to an immune mediated attack on normal HSC, but the evidence supporting this hypothesis is inconclusive...
June 18, 2018: PLoS Computational Biology
Tao Wang, Wu Tong, Chao Ye, Zhiqing Yu, Jing Chen, Fei Gao, Tongling Shan, Hai Yu, Liwei Li, Guoxin Li, Guangzhi Tong, Hao Zheng
Since late 2011, a pseudorabies virus (PRV) variant with increased virulence in old pigs had caused major disease outbreaks and great economic losses to the pig industry in China. The gene mutations that contributed to the increased virulence were unclear. To study the basis of the enhanced pathogenicity, an infectious bacterial artificial chromosome (BAC) clone consisting of the complete genome of the PRV variant was developed. Using homologous recombination and Cre/LoxP recombination, the recombinant virus rJS-2012-BAC carrying a BAC insertion downstream of the open reading frame (ORF) of gG was constructed...
June 9, 2018: Journal of Virological Methods
Jun-Gu Kang, Sung-Suck Oh, Young-Sun Jo, Jeong-Byoung Chae, Yoon-Kyoung Cho, Joon-Seok Chae
BACKGROUND: Severe fever with thrombocytopenia syndrome virus (SFTSV) is an emerging tick-borne virus in China, Japan, and Korea. Although the incidence of SFTS has been increasing annually since the first SFTSV case was reported in Korea, the prevalence of SFTSV in Korean livestock has not conducted. The purpose of this study was to investigate the prevalence of SFTSV in domesticated pigs (Sus scrofa domesticus) throughout Korea. METHODS: A total of 240 sera were collected from 30 domesticated pigs in eight provinces...
June 12, 2018: Vector Borne and Zoonotic Diseases
Reynaldo Ponce, Nancy León-Janampa, Robert H Gilman, Ruddy Liendo, Elisa Roncal, Sueline Luis, Stefany Quiñones-Garcia, Zach Silverstein, Hector H García, Armando Gonzales, Patricia Sheen, Mirko Zimic, Mónica Pajuelo
Cysticercosis is a worldwide parasitic disease of humans and pigs principally caused by infection with the larvae of the pork tapeworm Taenia solium. Through the use of the recently-made-available T. solium genome, we identified a gene within a novel 1448 bp ORF that theoretically encodes for a 433 amino acid-long protein and predicted to be an α-enolase closely related to enolases of other flatworms. Additional bioinformatic analyses revealed a putative plasminogen-binding region on this protein, suggesting a potential role for this protein in pathogenesis...
June 6, 2018: Experimental Parasitology
J Liu, W You, Z Xu, B Chen, Y Wang, T Shan
In the current study, we aimed to clone the full length cDNA of pCRTC3 gene and examine its expression pattern and function in intestinal epithelial cells. The full length cDNA sequence of pCRTC3 was 2173 bp (GenBank accession no. MF964215), with a 1,860-bp open reading frame encoding a 620-AA protein. Comparison of the deduced AA sequence with different species including human, mouse, rat, papio, cattle and rabbit showed 89% to 91.9% similarity. The pCRTC3 was highly expressed in small intestine and spleen, to a lesser degree in lung, liver and adipose tissue, and was expressed at a low but detectable level in skeletal muscle, kidney and heart...
May 23, 2018: Journal of Animal Science
Long Jin, Qing Guo, Guang-Lei Zhang, Xiao-Xu Xing, Mei-Fu Xuan, Qi-Rong Luo, Zhao-Bo Luo, Jun-Xia Wang, Xi-Jun Yin, Jin-Dan Kang
Epigenetic reprogramming and somatic cell nuclear transfer (SCNT) cloning efficiency were recently enhanced using histone deacetylase inhibitors (HDACis). In this study, we investigated the time effect of CI994, an HDACi, on the blastocyst formation rate, acetylation levels of H3K9 and H4K12, DNA methylation levels of anti-5-methylcytosine (5mC), and some mRNA expression of pluripotency-related genes in pig SCNT embryos. Treatment with 10 μM CI994 for 24 hours significantly improved the blastocyst formation rate of SCNT embryos in comparison with the untreated group (p < 0...
May 21, 2018: Cellular Reprogramming
Tingting Guo, Jinhu Huang, Changchao Huan, Fang He, Yujuan Zhang, Zohaib A Bhutto, Liping Wang
P-Glycoprotein (P-gp, Abcb1) plays a crucial role in drug disposition and functions by hydrolyzing ATP. However, little is known about the regulatory elements governing the transcription of the porcine Abcb1 gene. In this study, the transcription start site of the pig Abcb1 gene was identified by 5'-RACE. A 1.9-kb fragment of the 5'-flanking region of the Abcb1 gene was cloned from pig genomic DNA and sequenced. The region critical for its promoter activity was investigated via progressive deletions. Further, using mutation assays, two proximal Sp1 binding sites within the 5'-flanking region of Abcb1 were proven to be important cis- regulatory elements involved in regulating the constitutive expression of porcine Abcb1 ...
2018: Frontiers in Pharmacology
Lisa Niemann, Petra Müller, Jasmin Brauns, Rolf Nathaus, Franziska Schäkel, Kerstin Kipschull, Doris Höltig, Michael Wendt, Stefan Schwarz, Kristina Kadlec
The collaboration project VASIB aims at reducing the antibiotic consumption in pig production by integrating information from consulting expertise in clinical inspection, hygiene, epidemiology, microbiology and pharmacology. In this VASIB subproject, we investigated the antimicrobial susceptibility and relatedness of porcine respiratory tract pathogens. Bordetella bronchiseptica (n = 47), Pasteurella multocida (n = 18) and Streptococcus suis (n = 58) were obtained from weaner pigs at two farms. Antimicrobial susceptibility testing was performed by broth microdilution according to CLSI standards...
June 2018: Veterinary Microbiology
Katie Tungatt, Garry Dolton, Sophie B Morgan, Meriem Attaf, Anna Fuller, Thomas Whalley, Johanneke D Hemmink, Emily Porter, Barbara Szomolay, Maria Montoya, John A Hammond, John J Miles, David K Cole, Alain Townsend, Mick Bailey, Pierre J Rizkallah, Bryan Charleston, Elma Tchilian, Andrew K Sewell
There is increasing evidence that induction of local immune responses is a key component of effective vaccines. For respiratory pathogens, for example tuberculosis and influenza, aerosol delivery is being actively explored as a method to administer vaccine antigens. Current animal models used to study respiratory pathogens suffer from anatomical disparity with humans. The pig is a natural and important host of influenza viruses and is physiologically more comparable to humans than other animal models in terms of size, respiratory tract biology and volume...
May 2018: PLoS Pathogens
Meng Li, Xiaohong Guo, Pengfei Gao, Guoqing Cao, Zhimin Cheng, Wanfeng Zhang, Jianfeng Liu, Xiaojun Liu, Bugao Li
Objective: The aim of this study was to clone alternative splicing isoforms of pig MYNN, predict the structure and function of coding protein, and study temporal and spatial expression characteristics of each transcript. Methods: Alternative splice isoforms of MYNN were identified using RNA sequencing (RNA-seq) and cloning techniques. Quantitative real-time PCR (qPCR) was employed to detect expression patterns in 11 tissues of Large White (LW) and Mashen (MS) pigs, and to study developmental expression patterns in cerebellum (CE), stomach (ST) and longissimus dorsi (LD)...
April 25, 2018: Asian-Australasian Journal of Animal Sciences
Isabel Cornejo, Sandra Villanueva, Johanna Burgos, Karen I López-Cayuqueo, Régine Chambrey, Francisca Julio-Kalajzić, Neudo Buelvas, María I Niemeyer, Dulce Figueiras-Fierro, Peter D Brown, Francisco V Sepúlveda, L P Cid
Kir7.1 encoded by the Kcnj13 gene in the mouse is an inwardly rectifying K+ channel present in epithelia where it shares membrane localization with the Na+ /K+ -pump. Further investigations of the localisation and function of Kir7.1 would benefit from the availability of a knockout mouse, but perinatal mortality attributed to cleft palate in the neonate has thwarted this research. To facilitate localisation studies we now use CRISPR/Cas9 technology to generate a knock-in mouse, the Kir7.1-HA that expresses the channel tagged with a haemagglutinin (HA) epitope...
2018: Frontiers in Physiology
Shuchen Feng, Melinda Bootsma, Sandra L McLellan
The human microbiome contains many organisms that could potentially be used as indicators of human fecal pollution. Here we report the development of two novel human-associated genetic marker assays that target organisms within the family Lachnospiraceae Next-generation sequencing of the V6 region of the 16S rRNA gene from sewage and animal stool samples identified 40 human-associated marker candidates with a robust signal in sewage and low or no occurrence in nonhuman hosts. Two were chosen for quantitative PCR (qPCR) assay development using longer sequences (V2 to V9 regions) generated from clone libraries...
May 4, 2018: Applied and Environmental Microbiology
Molin Chen, Wei Tang, Xiuguo Hua
BACKGROUND: Porcine teschoviruses (PTVs) are small non-enveloped viruses with single-stranded, positive sense genomic RNA, belonging to the family Picornaviridae. Natural infections of teschoviruses are limited to pigs. RESULTS: In this study, a PTV HuN-1 was found that it could be proliferated in PK-15 cell, and it came from the pig fecal samples from Hunan province, in central China. The complete genome of the HuN-1 was amplified by RT-PCR and sequenced. The complete genome of HuN-1 isolate is 7098 nt, which shares the highest sequence identity (85...
April 27, 2018: BMC Veterinary Research
Min Jung Kim, Hyun Ju Oh, Yoo Bin Choi, Sanghoon Lee, Erif Maha Nugraha Setyawan, Seok Hee Lee, Seung Hoon Lee, Tai Young Hur, Byeong Chun Lee
This study was conducted to investigate whether the treatment of dog to pig interspecies somatic cell nuclear transfer (iSCNT) embryos with a histone deacetylase inhibitor, to improve nuclear reprogramming, can be applied to dog SCNT embryos. The dog to pig iSCNT embryos were cultured in fresh porcine zygote medium-5 (PZM-5) with 0, 1, or 10 µM suberoylanilide hydroxamic acid (SAHA) for 6 h, then transferred to PZM-5 without SAHA. Although there were no significant differences in cleavage rates, the rates of 5-8-cell stage embryo development were significantly higher in the 10 µM group (19...
April 26, 2018: Journal of Reproduction and Development
Aiwei Zhu, Tinglei Jiang, Tingsong Hu, Shijiang Mi, Zihan Zhao, Fuqiang Zhang, Jiang Feng, Quanshui Fan, Biao He, Changchun Tu
The family Circoviridae comprises a large group of small circular single-stranded DNA viruses with several members causing severe pig and poultry diseases. In recent years the number of new viruses within the family has had an explosive increase showing a high level of genetic diversity and a broad host range. In this report we describe two more circoviruses identified from bats in Yunnan and Heilongjiang provinces in China. Full genome sequencing has revealed that these bat associated circoviruses (bat ACV) should be classified as new species within the genus Circovirus based on the demarcation criteria of the International Committee on the Taxonomy of Viruses (ICTV)...
May 2, 2018: Virus Research
Z Yang, Y Hu, P Yuan, Y Yang, K Wang, L Y Xie, S L Huang, J Liu, L Ran, Z H Song
In the swine hepatitis E virus (HEV), open reading frame 2 (ORF2) is rich in antigenic determinants and neutralizing epitopes that could induce immune protection. We chose the Bac-to-Bac® Baculovirus Expression System to express fragments containing the critical neutralizing antigenic sites within the HEV ORF2 protein of pigs to obtain a recombinant baculovirus. The fragment of swine HEV ORF2 region (1198-1881bp) was cloned into vector pFastBacTM. A recombinant baculovirus, rBacmid-ORF2, was obtained after transposition and transfection...
March 2018: Polish Journal of Veterinary Sciences
Jiawen Wang, Shuai Bian, Meichun Liu, Xin Zhang, Siming Wang, Xueyuan Bai, Daqing Zhao, Yu Zhao
BST-2(tetherin/CD317/HM1.24) has been identified as a cellular antiviral factor that inhibits the release of a wide range of enveloped viruses from infected cells. Orthologs of BST-2 have been identified in several species including humans, monkeys, cows, sheep, pigs, and mice. In this study, we cloned the gene and characterized the protein of the BST-2 homolog from sika deer (Cervus nippon). cnBST-2 shares 37.8% and 74.2% identity with the BST-2 homologs from Homo sapiens and Ovis aries, respectively. The extracellular domain of cnBST-2 has two putative N-linked glycosylation sites and three potential dimerization sites...
June 30, 2018: Gene
Naresh L Selokar, Monika Saini, Prabhat Palta, Manmohan S Chauhan, Radhey S Manik, Suresh K Singla
Buffalo (Bubalus bubalis) is a major source of milk, meat, and draught power in many developing countries in Asia. Animal cloning holds a lot of potential for fast multiplication of elite buffaloes and conservation of their valuable germplasm. Although the progress of buffalo cloning has been slow in comparison to cattle or pig, several breakthroughs were reported in buffalo cloning such as the production of cloned calves from somatic cells isolated from over one-decade old frozen-thawed semen or from urine-derived cells...
April 2018: Cellular Reprogramming
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