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cloning pig

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https://www.readbyqxmd.com/read/29208927/establishment-of-a-strain-of-haemophilia-a-pigs-by-xenografting-of-foetal-testicular-tissue-from-neonatally-moribund-cloned-pigs
#1
Hiroyuki Kaneko, Kazuhiro Kikuchi, Michiko Nakai, Daiichiro Fuchimoto, Shunichi Suzuki, Shoichiro Sembon, Junko Noguchi, Akira Onishi
Grafting of testicular tissue into immunodeficient mice makes it possible to obtain functional sperm from immature donor animals that cannot be used for reproduction. We have developed a porcine model of human haemophilia A (haemophilia-A pigs) by nuclear transfer cloning from foetal fibroblasts after disruption of the X-linked coagulation factor VIII (F8) gene. Despite having a recessive condition, female F8+/- cloned pigs died of severe bleeding at an early age, as was the case for male F8-/Y cloned pigs, thus making it impossible to obtain progeny...
December 5, 2017: Scientific Reports
https://www.readbyqxmd.com/read/29184988/expression-and-characterization-of-a-recombinant-porcinized-antibody-against-the-e2-protein-of-classical-swine-fever-virus
#2
Shucheng Chen, Su Li, Huimin Sun, Yongfeng Li, Shengwei Ji, Kun Song, Lingkai Zhang, Yuzi Luo, Yuan Sun, Jifei Ma, Pinghuang Liu, Hua-Ji Qiu
Classical swine fever virus (CSFV) is the causative agent of classical swine fever (CSF), a highly contagious and economically important disease of pigs. The envelope glycoprotein E2 of CSFV is the major antigen that induces neutralizing antibodies and confers protection against CSFV infections. Previously, we developed a murine monoclonal antibody (MAb), HQ06, against the E2 protein of CSFV. To produce the antibody conveniently and stably, the genes coding for the variable regions of the heavy and light chains of HQ06 and constant region genes from the swine antibody were fused and cloned into lentiviral expression vectors to express a recombinant porcinized MAb (rHQ06Sw) in mammalian cells...
November 28, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29180167/characterization-of-equine-gst-a3-3-as-a-steroid-isomerase
#3
Helena Lindström, Shawna M Peer, Nancy H Ing, Bengt Mannervik
Glutathione transferases (GSTs) comprise a superfamily of enzymes prominently involved in detoxication by making toxic electrophiles more polar and therefore more easily excretable. However some GSTs have developed alternative functions. Thus, a member of the Alpha class GSTs in pig and human tissues is involved in steroid hormone biosynthesis, catalyzing the obligatory double-bond isomerization of Δ5-androstene-3,17-dione to Δ4-androstene-3,17-dione and of Δ5-pregnene-3,20-dione to Δ4-pregnene-3,20-dione on the biosynthetic pathways to testosterone and progesterone...
November 24, 2017: Journal of Steroid Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/29177583/identification-and-functional-analysis-of-the-novel-orf6-protein-of-porcine-circovirus-type-2-in-vitro
#4
Dongge Li, Jing Wang, Shangen Xu, Shanxia Cai, Chaojie Ao, Liurong Fang, Shaobo Xiao, Huanchun Chen, Yunbo Jiang
In the present study, the function of a novel ORF6 gene in the PCV2 genome was determined and functionally analyzed in vitro. ORF6 expression was demonstrated by indirect immunofluorescence in PCV2-infected cells. The antibody against ORF6 was detected in PCV2-infected pigs. The start codon of ORF6 was mutated and an infectious clone was used to create an ORF6-deficient mutant virus. Viral DNA replication curves and immunofluorescence analysis indicated that ORF6 is unnecessary for viral replication and ORF6 deletion reduces viral DNA replication in PK-15 cells...
November 25, 2017: Veterinary Research Communications
https://www.readbyqxmd.com/read/29154473/influences-of-quorum-quenching-probiotic-bacteria-on-the-gut-microbial-community-and-immune-function-in-weaning-pigs
#5
Jonggun Kim, Jaepil Kim, Younghoon Kim, Sangnam Oh, Minho Song, Jee Hwan Choe, Kwang-Youn Whang, Kwang Hyun Kim, Sejong Oh
The aim of this study is to investigate the dynamic gut microbial diversity in weaning swine after administering feed supplemented with probiotic bacteria that specifically inhibit the activity of quorum molecules. Initially, the universal quorum molecule autoinducer-2 (AI-2) bioassay results indicated that AI-2 activity was profoundly inhibited in enterohemorrhagic Escherichia coli (EHEC) O157:H7 in the presence of Lactobacillus acidophilus strain 30SC cell extract, although the growth of EHEC was not affected...
November 20, 2017: Animal Science Journal, Nihon Chikusan Gakkaihō
https://www.readbyqxmd.com/read/29150045/spectrum-of-benzo-a-pyrene-induced-mutations-in-the-pig-a-gene-of-l5178ytk-cells-identified-with-next-generation-sequencing
#6
Javier Revollo, Yiying Wang, Page McKinzie, Azra Dad, Mason Pearce, Robert H Heflich, Vasily N Dobrovolsky
We used Sanger sequencing and next generation sequencing (NGS) for analysis of mutations in the endogenous X-linked Pig-a gene of clonally expanded L5178YTk(+/-) cells. The clones developed from single cells that were sorted on a flow cytometer based upon the expression pattern of the GPI-anchored marker, CD90, on their surface. CD90-deficient and CD90-proficient cells were sorted from untreated cultures and CD90-deficient cells were sorted from cultures treated with benzo[a]pyrene (B[a]P). Pig-a mutations were identified in all clones developed from CD90-deficient cells; no Pig-a mutations were found in clones of CD90-proficient cells...
December 2017: Mutation Research
https://www.readbyqxmd.com/read/29121641/identification-of-a-t-3-4-p1-3-q1-5-translocation-breakpoint-in-pigs-using-somatic-cell-hybrid-mapping-and-high-resolution-mate-pair-sequencing
#7
Katia Fève, Sylvain Foissac, Alain Pinton, Florence Mompart, Diane Esquerré, Thomas Faraut, Martine Yerle, Juliette Riquet
Reciprocal translocations are the most frequently occurring constitutional structural rearrangements in mammalian genomes. In phenotypically normal pigs, an incidence of 1/200 is estimated for such rearrangements. Even if constitutional translocations do not necessarily induce defects and diseases, they are responsible for significant economic losses in domestic animals due to reproduction failures. Over the last 30 years, advances in molecular and cytogenetic technologies have led to major improvements in the resolution of the characterization of translocation events...
2017: PloS One
https://www.readbyqxmd.com/read/29115020/glycosylphosphatidylinositol-gpi-anchored-protein-deficiency-serves-as-a-reliable-reporter-of-pig-a-gene-mutation-support-from-an-in-vitro-assay-based-on-l5178y-tk-cells-and-the-cd90-2-antigen
#8
Jeffrey C Bemis, Svetlana L Avlasevich, Carson Labash, Page McKinzie, Javier Revollo, Vasily N Dobrovolsky, Stephen D Dertinger
Lack of cell surface glycosylphosphatidylinositol (GPI)-anchored protein(s) has been used as a reporter of Pig-a gene mutation in several model systems. As an extension of this work, our laboratory initiated development of an in vitro mutation assay based on the flow cytometric assessment of CD90.2 expression on the cell surface of the mouse lymphoma cell line L5178Y/Tk(+/-) . Cells were exposed to mutagenic and nonmutagenic compounds for 24 hr followed by washout and incubation for an additional 7 days. Following this mutant manifestation time, cells were labeled with fluorescent antibodies against CD90...
November 8, 2017: Environmental and Molecular Mutagenesis
https://www.readbyqxmd.com/read/29100547/efficient-generation-of-p53-biallelic-knockout-diannan-miniature-pigs-via-talens-and-somatic-cell-nuclear-transfer
#9
Youfeng Shen, Kaixiang Xu, Zaimei Yuan, Jianxiong Guo, Heng Zhao, Xuezeng Zhang, Lu Zhao, Yubo Qing, Honghui Li, Weirong Pan, Baoyu Jia, Hong-Ye Zhao, Hong-Jiang Wei
BACKGROUND: Pigs have many features that make them attractive as biomedical models for various diseases, including cancer. P53 is an important tumor suppressor gene that exerts a central role in protecting cells from oncogenic transformation and is mutated in a large number of human cancers. P53 mutations occur in almost every type of tumor and in over 50% of all tumors. In a recent publication, pigs with a mutated P53 gene were generated that resulted in lymphoma and renal and osteogenic tumors...
November 3, 2017: Journal of Translational Medicine
https://www.readbyqxmd.com/read/29098723/establishing-a-novel-pig-a-gene-mutation-assay-in-l5178ytk-mouse-lymphoma-cells
#10
Yiying Wang, Javier Revollo, Page McKinzie, Mason G Pearce, Azra Dad, Berran Yucesoy, Hans Rosenfeldt, Robert H Heflich, Vasily N Dobrovolsky
The X-linked Pig-a gene encodes an enzyme required for the biosynthesis of glycosyl phosphatidylinositol (GPI) anchors. Pig-a mutant cells fail to synthesize GPI and to express GPI-anchored protein markers (e.g., CD90) on their surface. Marker deficiency serves as a phenotypic indicator of Pig-a mutation in various in vivo assays. Here, we describe an in vitro Pig-a mutation assay in L5178YTk(+/-) mouse lymphoma cells, in which mutant-phenotype cells are measured by flow cytometry using a fluorescent anti-CD90 antibody...
November 2, 2017: Environmental and Molecular Mutagenesis
https://www.readbyqxmd.com/read/29080625/diversity-of-livestock-associated-methicillin-resistant-staphylococcus-aureus
#11
Henok A Tegegne, Ivana Koláčková, Renata Karpíšková
OBJECTIVE: To evaluate the diversity and molecular characteristics of livestock associated methicillin-resistant Staphylococcus aureus in livestock animals, food of animal origin and the environment in the Czech Republic. METHODS: After having been primarily enriched in buffered peptone water, the samples were cultured on Baird-Parker agar. Presumptive colonies were sub-cultured to blood agar and assessed morphologically. Furthermore, presumptive Staphylococcus aureus colonies were confirmed by MALDI-TOF...
September 2017: Asian Pacific Journal of Tropical Medicine
https://www.readbyqxmd.com/read/29069793/imprinting-disorder-in-donor-cells-is-detrimental-to-the-development-of-cloned-embryos-in-pigs
#12
Xuexiong Song, Fangzheng Li, Zhongling Jiang, Yueping Sun, Huatao Li, Shansong Gao, Liping Zhang, Binghua Xue, Guimin Zhao, Jingyu Li, Zhonghua Liu, Hongbin He, Yanjun Huan
Imprinting disorder during somatic cell nuclear transfer usually leads to the abnormality of cloned animals and low cloning efficiency. However, little is known about the role of donor cell imprinting in the development of cloned embryos. Here, we demonstrated that the imprinting (H19/Igf2) in porcine fetus fibroblasts derived from the morphologically abnormal cloned fetuses (the abnormal imprinting group) was more hypomethylated, and accordingly, significantly higher H19 transcription and lower Igf2 expression occurred in comparison with those in fibroblasts derived from morphologically normal cloned fetuses (the normal imprinting group) or donor fetus fibroblasts (the control group)...
September 22, 2017: Oncotarget
https://www.readbyqxmd.com/read/29069323/salmonella-enterica-serotype-4-5-12-i-in-swine-in-the-united-states-midwest-an-emerging-multidrug-resistant-clone
#13
Ehud Elnekave, Samuel Hong, Alison E Mather, Dave Boxrud, Angela J Taylor, Victoria Lappi, Timothy J Johnson, Fabio Vannucci, Peter Davies, Craig Hedberg, Andres Perez, Julio Alvarez
Background: Salmonella 4,[5],12:i:-, a worldwide emerging pathogen that causes many foodborne outbreaks mostly attributed to pig and pig products, is expanding in the U.S. Methods: Whole genome sequencing was applied to conduct multiple comparisons of 659 S. 4,[5],12:i:- and 325 S. Typhimurium from different sources and locations (i.e. U.S. and Europe) to assess their genetic heterogeneity, with a focus on strains recovered from swine in the U.S. Midwest. In addition, presence of resistance genes and other virulence factors was detected and the antimicrobial resistance phenotype of 50 and 22 isolates of livestock and human origin, respectively, was determined...
October 23, 2017: Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America
https://www.readbyqxmd.com/read/29059459/molecular-cloning-and-tissue-distribution-of-a-novel-marmoset-abc-transporter
#14
Shotaro Uehara, Yasuhiro Uno, Takashi Inoue, Erika Sasaki, Hiroshi Yamazaki
Common marmosets (Callithrix jacchus) have been recognized as a useful small non-human primate model in preclinical testing for drug development. In this study, a cDNA of novel ATP-dependent efflux transporter ABCB1 was cloned from marmoset liver tissue. Marmoset ABCB1 cDNA encodes a protein of 1,279 amino acid residues (MW = 141.4 kDa) containing characteristic regions of an ATP-binding cassette (ABC) protein, two hydrophobic transmembrane regions, and two cytoplasmic nucleotide-binding regions, similar to human ABCB1...
October 23, 2017: Biopharmaceutics & Drug Disposition
https://www.readbyqxmd.com/read/29058487/lc3-dependent-autophagy-in-pig-2-cell-cloned-embryos-could-influence-the-degradation-of-maternal-mrna-and-the-regulation-of-epigenetic-modification
#15
Daming Chi, Yaqiong Zeng, Mingzhu Xu, Linan Si, Xiao Qu, Honglin Liu, Juan Li
In this study, the distribution as well as the effect of autophagy on reprogramming in pig cloned embryos were observed immediately after somatic cell nuclear transfer. Results showed that the LC3 was at the highest level in cloned embryos at 2-cell stage, and it decreased with the development from 2-cell stage to blastocyst. Different to cloned embryos, the intensity of LC3 in parthenogenetic activation (PA) embryos was at the highest level at 4-cell stage. A markedly higher level of Bmp15, H1foo, and Dppa3 was shown in cloned embryos at 2-cell stage (p < 0...
October 23, 2017: Cellular Reprogramming
https://www.readbyqxmd.com/read/29048488/clinical-and-molecular-features-of-mdr-livestock-associated-mrsa-st9-with-staphylococcal-cassette-chromosome-mecxii-in-humans
#16
Chih-Jung Chen, Tsai-Ling Yang Lauderdale, Chin-Te Lu, Yu-Yu Chuang, Chien-Chang Yang, Ting-Shu Wu, Chun-Yi Lee, Min-Chi Lu, Wen-Chien Ko, Yhu-Chering Huang
Objectives: Clonal complex (CC) 9 is a prevalent livestock-associated (LA) MRSA clone in Asia whose pathogenicity in humans remains unknown. Methods: In 2012, we identified a patient with CC9-MRSA infection linked to livestock. After screening 3328 clinical MRSA isolates from a national database, eight isolates (0.24%) collected between 1998 and 2012 were further confirmed to be of CC9. The detailed molecular features of the nine human CC9 strains and phylogenetic relatedness to animal CC9 strains were characterized with WGS...
October 18, 2017: Journal of Antimicrobial Chemotherapy
https://www.readbyqxmd.com/read/29047027/a-possible-mechanism-for-low-affinity-of-silkworm-na-k-atpase-for-k
#17
Haruo Homareda, Masahiro Otsu, Sachiko Yamamoto, Makoto Ushimaru, Sayaka Ito, Toshiyuki Fukutomi, Taeho Jo, Yoshinobu Eishi, Yukichi Hara
The affinity for K(+) of silkworm nerve Na(+)/K(+)-ATPase is markedly lower than that of mammalian Na(+)/K(+)-ATPase (Homareda 2010). In order to obtain clues on the molecular basis of the difference in K(+) affinities, we cloned cDNAs of silkworm (Bombyx mori) nerve Na(+)/K(+)-ATPase α and β subunits, and analyzed the deduced amino acid sequences. The molecular masses of the α and β subunits were presumed to be 111.5 kDa with ten transmembrane segments and 37.7 kDa with a single transmembrane segment, respectively...
October 19, 2017: Journal of Bioenergetics and Biomembranes
https://www.readbyqxmd.com/read/29040878/c-phycocyanin-supplementation-during-in%C3%A2-vitro-maturation-enhances-pre-implantation-developmental-competence-of-parthenogenetic-and-cloned-embryos-in-pigs
#18
Shuang Liang, Jing Guo, Yong Xun Jin, Bao Yuan, Jia-Bao Zhang, Nam-Hyung Kim
C-Phycocyanin (C-PC), a protein from green microalgae, has been suggested to possess various biological activities, including antioxidant and free radical scavenging properties. The aim of the current study was to explore the effects of C-PC on the maturation of porcine oocytes and subsequent developmental competence after parthenogenetic activation and somatic cell nuclear transfer (SCNT) as well as the underlying mechanisms. There was no significant improvement in nuclear maturation rates between the control and C-PC supplementation groups (1, 3, 5, 10 μg/mL)...
September 8, 2017: Theriogenology
https://www.readbyqxmd.com/read/29030661/identification-of-the-major-histocompatibility-complex-class-ii-dm-and-do-alleles-in-a-cohort-of-northern-pig-tailed-macaques-macaca-leonina
#19
Xiao-Dong Lian, Xi-He Zhang, Zheng-Xi Dai, Yong-Tang Zheng
The northern pig-tailed macaque (Macaca leonina) has been considered as an independent species from the pig-tailed macaque group. We have previously reported that this species macaque has the potential to be a useful animal model in HIV/AIDS pathogenesis and vaccine studies due to its susceptibility to HIV-1. To develop this animal into a potential HIV/AIDS model, we have studied the classical MHC genes of this animal. In this study, the non-classical MHC genes Malo-DM and Malo-DO alleles were first characterized by sequencing and cloning in 12 unrelated northern pig-tailed macaques...
October 13, 2017: Immunogenetics
https://www.readbyqxmd.com/read/28993973/crispr-cas9-mediated-knockout-of-myostatin-in-chinese-indigenous-erhualian-pigs
#20
Kankan Wang, Xiaochun Tang, Zicong Xie, Xiaodong Zou, Mengjing Li, Hongming Yuan, Nannan Guo, Hongsheng Ouyang, Huping Jiao, Daxin Pang
CRISPR/Cas9 has emerged as one of the most popular genome editing tools due to its simple design and high efficiency in multiple species. Myostatin (MSTN) negatively regulates skeletal muscle growth and mutations in myostatin cause double-muscled phenotype in various animals. Here, we generated myostatin mutation in Erhualian pigs using a combination of CRISPR/Cas9 and somatic cell nuclear transfer. The protein level of myostatin precursor decreased dramatically in mutant cloned piglets. Unlike myostatin knockout Landrace, which often encountered health issues and died shortly after birth, Erhualian pigs harboring homozygous mutations were viable...
October 9, 2017: Transgenic Research
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