Roslin Institute Research Articles

돼지 브랜드 식별 및 원산지 추적에 활용 가능한 Microsatellite Marker Set의 확립

돼지 원산지 추적 및 브랜드육 식별을 위한 MS marker set를 확립하기 위해 EU의 EID+DNA DNA Tracing 연구 project, 국제동물유전학회 및 Roslin 연구소 등에서 제안한 돼지의 다형성 분석에 사용되는 MS marker 중 17종을 선발하여 다형성지수, F-통계량, 동일개체 출현확률 및 친자감별확률 등을 MSA, CERVUS, FSTAT, GENEPOP 및 API-CALC 프로그램 등을 연계적으로 활용하여 추정하였다. 다형성지수 추정치를 1차 선발요인으로 하고, 각 MS marker의 PCR 결과물의 크기 및 분석 비용의 경제성을 추가로 감안하여 최종 13종의 MS marker (SW936, SW951, SW787, S00090, S0026, SW122, SW857, S0005, SW72, S0155, S0225, SW24 and SW632)와 성감별을 위한 2개의 성감별 primer로 조합된 하나의 Multiplexing PCR 시스템을 확립하였다. 이를 이용해 돼지 사육환경을 무작위 교배집단(PI), 반형매 교배집단 그리고 전형매 교배집단으로 가정하여 동일개체 출현확률을 분석한 결과 <TEX>$2.47\times10^{-18}$</TEX>, <TEX>$6.39\times10^{-13}$</TEX> 그리고 <TEX>$1.08\times10^{-8}$</TEX>로 나타나 브랜드 식별 및 원산지 추적에 적합할 것으로 사료된다. Seventeen porcine microsatellite (MS) markers recommended by the EID+DNA Tracing EU project, ISAG and Roslin institute were selected for the use in porcine individual and brand identification. The MSA, CERVUS, FSTAT, GENEPOP and API-CALC programs were applied for calculating heterozygosity indices. By considering the hetreozygosity value and PCR product size of each marker, we established a MS marker set composed of 13 MS markers (SW936, SW951, SW787, S00090, S0026, SW122, SW857, S0005, SW72, S0155, S0225, SW24 and SW632) and two sexing markers. The expected probability of identity among genotypes of random individuals (PI), probability of identity among genotypes from random half sibs (<TEX>$PI_{half-sibs}$</TEX>) and among genotypes of random individuals, probability of identity among genotypes from random sibs(<TEX>$PI_{sibs}$</TEX>) were estimated as <TEX>$2.47\times10^{-18}$</TEX>, <TEX>$6.39\times10^{-13}$</TEX> and <TEX>$1.08\times10^{-8}$</TEX>, respectively. The results indicate that the established marker set can provide a sufficient discriminating power in both individual and parentage identification for the commercial pigs produced in Korea.

Identification of Adjacent Binding Sites for the YY1 and E4BP4 Transcription Factors in the Ovine PrP (Prion) Gene Promoter

The PrP gene encodes the cellular isoform of the prion protein (PrP(c)) which has been shown to be crucial to the development of transmissible spongiform encephalopathies (TSEs). PrP knock-out mice, which do not express endogenous PrP(c), exhibit resistance to TSE disease. The regulation of PrP gene expression represents, therefore, a crucial factor in the development of TSEs. Two sequence motifs in the PrP promoter (positions -287 to -263 from transcriptional start) were previously reported as being highly conserved, and it was suggested that they represent binding sites for as yet unidentified transcription factors. To test this hypothesis, binding of nuclear proteins was analyzed by electrophoretic mobility shift assays using ovine or murine cells and tissues with radiolabeled DNA probes containing the conserved motif sequences. Specific binding was observed to both motifs, and polymorphic variants of these motifs exhibited differential binding. Two proteins bound to these motifs were identified as the Yin Yang 1 (YY1) (motif 1) and E4BP4 (motif 2) transcription factors. Functional promoter analysis of four different promoter variants revealed that motif 1 (YY1) was associated with inhibitory activity in the context of the PrP promoter, whereas motif 2 (E4BP4) was linked to a slight enhancing activity. This represents the first demonstration of binding of nuclear factors to two highly conserved DNA sequence motifs within mammalian PrP promoters. The action of these factors on the PrP promoter is haplotype-specific, leading us to propose that the prion protein expression pattern and, with it, the distribution of TSE infectivity may be associated with PrP promoter genotype.

The 2008 Dolph Adams Award and the State of the Journal of Leukocyte Biology

The 2008 Dolph Adams Award and the State of the <i>Journal of Leukocyte Biology</i>

Induced Pluripotent Stem Cells: a New Hope or a New Controversy?

It seems unbelievable but it is true – cells within an organism are genetically almost identical and yet they are as dissimilar structurally and functionally as the human mind can conceive. The neurons which conduct the nerve impulses, the villi which line the gastrointestinal tract, the retinal cells which are responsible for our vision, and the immune cells which act as the body’s sentinel have hardly anything in common between them. All these highly specialized cells are said to be terminally differentiated - they have lost their ability to become something else. Therefore, it was heralded as a significant breakthrough in science, when more than a decade ago, Ian Wilmut and colleagues at the Roslin Institute in Scotland demonstrated that a mature differentiated somatic cell taken from an adult mammal could be reprogrammed to give rise to an entire animal, which came to be known to the entire world as Dolly the sheep (Campbell 1996, 64).

‘Crook’ pipettes: embryonic emigrations from agriculture to reproductive biomedicine

While cloning, stem cells, and regenerative medicine are often imagined in a futurial idiom—as expectations, hype, hope and promises—this article approaches the remaking of genealogy in such contexts from a historical route. Through a series of somewhat disparate historical connections linking Australian sheep to the development of clinical IVF and the cloning of Dolly at the Roslin Institute in Scotland in 1996, this article explores the linkages through which agriculture, embryology, and reproductive biomedicine are thickly intertwined. Key to this examination is not only the history of experimental sheep breeding, and its somewhat unexpectedly genealogical connections to (Australian) national identity (‘wool in the veins’), but also the re-emergence of a distinctive frontier ethos in the context of assisted conception, and later human embryonic stem cell derivation. I have set this scene of genealogical interconnection against the criss-crossing traffic between Britain and Australia, and the wool trade, to emphasise the importance of global, as well as local, connections in the bloodlines of animals such as Dolly. In sum, this article examines the idea of the ‘biological frontier’ by exploring its histories as a means to offset the assumption that this frequently encountered idiom describes a future that is, or must be, by definition, unknown and unknowable.

Human cloning a decade after Dolly

Ten years ago this month, Ian Wilmut and colleagues from the Roslin Institute in Scotland published an article in Nature unceremoniously entitled: “Viable offspring derived from fetal and adult mammalian cells.” This was, of course, the publication that announced the birth of Dolly the cloned

Physiological genomics special issue on animal functional genomics

this special issue of Physiological Genomics highlights 12 papers that reflect presentations made by 110 international delegates of the 2nd International Symposium on Animal Functional Genomics (2nd ISAFG) held May 16th-19th, 2006, at Michigan State University (East Lansing, MI). This was the second

Insulin-like Growth Factor-binding Protein-5 Activates Plasminogen by Interaction with Tissue Plasminogen Activator, Independently of Its Ability to Bind to Plasminogen Activator Inhibitor-1, Insulin-like Growth Factor-I, or Heparin

Transgenic mice expressing IGFBP-5 in the mammary gland exhibit increased cell death and plasmin generation. Because IGFBP-5 has been reported to bind to plasminogen activator inhibitor-1 (PAI-1), we determined the effects of this interaction in HC11 cells. PAI-1 prevented plasmin generation from plasminogen and inhibited cleavage of focal adhesions, expression of caspase 3, and cell death. IGFBP-5 could in turn prevent the effects of PAI-1. IGFBP-5 mutants with reduced affinity for IGF-I (N-term) or deficient in heparin binding (HEP- and C-term E and F) were also effective. This was surprising because IGFBP-5 reportedly interacts with PAI-1 via its heparin-binding domain. Biosensor analysis confirmed that, although wild-type IGFBP-5 and N-term both bound to PAI-1, the C-term E had greatly decreased interaction with PAI-1. This suggests that IGFBP-5 does not antagonize the actions of PAI-1 by a direct molecular interaction. In a cell-free system, using tissue plasminogen activator (tPA) and urokinase plasminogen activator (uPA) to activate plasminogen, PAI-1 inhibited plasmin generation induced by both activators, whereas IGFBP-5 prevented the effects of PAI-1 on tPA but not uPA. Furthermore, we noted that IGFBP-5 activated plasminogen to a greater extent than could be explained solely by inhibition of PAI-1, suggesting that IGFBP-5 could directly activate tPA. Indeed, IGFBP-5 and the C-term E and F were all able to enhance the activity of tPA but not uPA. These data demonstrate that IGFBP-5 can enhance the activity of tPA and that this can result in cell death induced by cleavage of focal adhesions. Thus IGFBP-5 can induce cell death by both sequestering IGF-I and enhancing plasmin generation.

Parental drift

Parental drift

A walk from the wild side: The genetics of domestication of livestock and crops

The phenotypic variation found in domesticated plants and animals is striking, so much so that Darwin used it to illustrate the power of selection to effect change. Recent developments in genomics technologies are leading to dramatic progress in elucidating the genetic changes that occur during domestication. The Genetics Society Autumn Meeting on the genetics of domestication took place in November 2004 at the Royal Society in London, and was organised by Helen Sang (Roslin Institute, UK) and Jonathan Jones (John Innes Centre, UK). The meeting brought together many of the leading researchers on livestock and crop domestication and provided a timely and exciting account of recent progress in the field.

Development of a fully anaesthetised porcine model of acetaminophen-induced acute liver failure: Philip N Newsome, Neil C Henderson, Steven Keatch, Leonard J Nelson, Celine Filippi, Chris Bellamy, Eddie Clutton, University of Edinburgh, Edinburgh United Kingdom; Tim King, Roslin Institute, Edinburgh United Kingdom; Alistair Lee, Royal Infirmary of Edinburgh, Edinburgh United Kingdom; Peter C Hayes, John N Plevris, University of Edinburgh,

Development of a fully anaesthetised porcine model of acetaminophen-induced acute liver failure: Philip N Newsome, Neil C Henderson, Steven Keatch, Leonard J Nelson, Celine Filippi, Chris Bellamy, Eddie Clutton, University of Edinburgh, Edinburgh United Kingdom; Tim King, Roslin Institute, Edinburgh United Kingdom; Alistair Lee, Royal Infirmary of Edinburgh, Edinburgh United Kingdom; Peter C Hayes, John N Plevris, University of Edinburgh,

Mapping of the bovine homologue of the human chromosome 14 open reading frame 4 (C14orf4) gene to BTA10q36

Animal GeneticsVolume 35, Issue 6 p. 498-499 Mapping of the bovine homologue of the human chromosome 14 open reading frame 4 (C14orf4) gene to BTA10q36 C. Drögemüller, C. Drögemüller Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorH. Kuiper, H. Kuiper Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorA. Spötter, A. Spötter Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorJ. L. Williams, J. L. Williams Roslin Institute (Edinburgh), Roslin, Midlothian, EH25 9PS, UKSearch for more papers by this authorO. Distl, O. Distl Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this author C. Drögemüller, C. Drögemüller Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorH. Kuiper, H. Kuiper Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorA. Spötter, A. Spötter Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorJ. L. Williams, J. L. Williams Roslin Institute (Edinburgh), Roslin, Midlothian, EH25 9PS, UKSearch for more papers by this authorO. Distl, O. Distl Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this author First published: 29 November 2004 https://doi.org/10.1111/j.1365-2052.2004.01211.xCitations: 1 C. Drögemüller ([email protected]) Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article.Citing Literature Volume35, Issue6December 2004Pages 498-499 RelatedInformation

High road to Scotland

The creation of Dolly the sheep by nuclear transfer at the Roslin Institute focused the world's attention on Scotland, and private funding and investment in public infrastructure may sustain this interest. This is especially true at the three biggest biomedical hubs: Edinburgh, Glasgow and Dundee, which are adding substantial life-science facilities over the next few years.

Assignment of the PRPH gene to bovine chromosome 5q1.4 by FISH and confirmation by RH mapping

Animal GeneticsVolume 35, Issue 6 p. 477-478 Assignment of the PRPH gene to bovine chromosome 5q1.4 by FISH and confirmation by RH mapping S. Mömke, S. Mömke Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorH. Kuiper, H. Kuiper Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorA. Spötter, A. Spötter Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorC. Drögemüller, C. Drögemüller Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorJ. L. Williams, J. L. Williams Roslin Institute (Edinburgh), Roslin, Midlothian, UKSearch for more papers by this authorO. Distl, O. Distl Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this author S. Mömke, S. Mömke Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorH. Kuiper, H. Kuiper Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorA. Spötter, A. Spötter Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorC. Drögemüller, C. Drögemüller Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this authorJ. L. Williams, J. L. Williams Roslin Institute (Edinburgh), Roslin, Midlothian, UKSearch for more papers by this authorO. Distl, O. Distl Institute for Animal Breeding and Genetics, University of Veterinary Medicine, Hannover, GermanySearch for more papers by this author First published: 29 October 2004 https://doi.org/10.1111/j.1365-2052.2004.01199.xCitations: 1 O. Distl (ottmar.distl@tiho-hannover.de) Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article.Citing Literature Volume35, Issue6December 2004Pages 477-478 RelatedInformation

Media Coverage of Cloning: A Study of Media Content, Production and Reception

This paper presents the results of empirical research that analyzed UK news media coverage of cloning. More specifically, it describes how quantitative and qualitative methods were used to examine the production, content and reception of newspaper and television news coverage of cloning. The paper documents the results of a systematic analysis of two years of media content (1996 and 1997), a period that includes the announcement that a Finn Dorset sheep (Dolly) had been cloned from a somatic cell. Interviews with media professionals and the Roslin Institute examined the processes of mediation involved in producing this coverage. A reception analysis, which investigated the significance of this coverage in informing respondents’ views about cloning, showed that these respondents were particularly influenced by coverage of Dolly the sheep. In conclusion, the paper considers how media coverage of cloning might influence the construction of scientific citizenship.

Preliminary linkage map of the Turkey (Meleagris gallopavo) based on microsatellite markers

The turkey is an agriculturally important species for which, until now, there is no published genetic linkage map based on microsatellite markers--still the markers most used in the chicken and other farm animals. In order to increase the number of markers on a turkey genetic linkage map we decided to map new microsatellite sequences obtained from a GT-enriched turkey genomic library. In different chicken populations more than 35-55% of microsatellites are polymorphic. In the turkey populations tested here, 43% of all turkey primers tested were found to be polymorphic, in both commercial and wild type turkeys. Twenty linkage groups (including the Z chromosome) containing 74 markers have been established, along with 37 other unassigned markers. This map will lay the foundations for further genetic mapping and the identification of genes and quantitative trait loci in this economically important species. Genome comparisons, based on genetic maps, with related species such as the chicken would then also be possible. All primer information, polymerase chain reaction (PCR) conditions, allele sizes and genetic linkage maps can be viewed at http://roslin.thearkdb.org/. The DNA is also available on request through the Roslin Institute.

Dolly Goes to Greener Pastures

Dolly the sheep, the first mammal cloned from an adult cell, died last week. Her caretakers at the Roslin Institute in Scotland euthanized the 6-year-old sheep on 14 February after diagnosing an incurable lung disease, pulmonary adenomatosis or Jaagsiekte. Her death seems unconnected to her unconventional origin, as the virus that causes the disease has infected both cloned and normal sheep at the institute.

Physical and genetic mapping of the porcine colipase gene ( CLPS ).

Animal GeneticsVolume 33, Issue 5 p. 377-377 Physical and genetic mapping of the porcine colipase gene (CLPS) J. F. Brown, J. F. Brown Roslin Institute (Edinburgh), Roslin, Midlothian, UKSearch for more papers by this authorA. L. Archibald, A. L. Archibald Roslin Institute (Edinburgh), Roslin, Midlothian, UKSearch for more papers by this author J. F. Brown, J. F. Brown Roslin Institute (Edinburgh), Roslin, Midlothian, UKSearch for more papers by this authorA. L. Archibald, A. L. Archibald Roslin Institute (Edinburgh), Roslin, Midlothian, UKSearch for more papers by this author First published: 30 September 2002 https://doi.org/10.1046/j.1365-2052.2002.00896_1.xCitations: 1Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Citing Literature Volume33, Issue5October 2002Pages 377-377 RelatedInformation

It's good to talk. Media training workshops are aiming to break down the barriers between scientists and journalists.

Soon after the birth of Dolly the Sheep, Jim McWhir from the Roslin Institute in Edinburgh arrived in Spain, where he was ushered into an unscheduled press conference. After some difficult questions about cloning, he was asked about a report that had appeared that morning in the Spanish press in which an American scientist claimed that, in the near future, men would be able to get pregnant. ‘I replied that I was not familiar with the work but that I wasn't about to lose any sleep worrying about getting pregnant,’ said McWhir, describing the experience. ‘So after being grilled about lots of serious issues, all that appeared on the Spanish news that evening was 10 seconds of me saying that I wasn't worried about getting pregnant.’ When science hits the headlines in instances like these, the wide gulf between scientists and the media is cringingly apparent. The problems start when a science story sprouts legs and walks from the laboratory to the news desk to become a hastily constructed article more about politics, health or ethics. From many scientists’ point of view, the publicity resulting from such coverage can be damaging, with the release of unpublished results, equal weight given to the scientific consensus and a minority opinion, and the word ‘breakthrough’ appearing far more frequently than it deserves. This very loss of control over the outcome of media encounters leads many scientists to be reluctant to even speak to journalists in the belief that they are exploitative, manipulative and—the ultimate sin—inaccurate in their representation of the facts. ![][1] Clearly, the media places a different emphasis on current events, but Pallab Ghosh, a science correspondent of the BBC and Chairman of the Association of British Science Writers, believes that this should be viewed more positively. ‘It's not a perfect system, but … [1]: /embed/graphic-1.gif

Stem Cells, Embryos and Cloning—Unravelling the Ethics of a Knotty Debate

The cloning of Dolly the sheep from somatic cells in 1996, the isolation of human embryonic stem cells in 1998, and the publication of the first drafts of the human genome in 2000 represent frontiers of remarkable new realms into which the biosciences are seeking to go. Each has been heralded by promises of untold benefits to be found in the new lands. Alongside the frequent rhetoric of progress and conquest, however, have also come many expressions of concern that some of the country ahead presents difficult ethical and social problems, which require treading carefully and some regions that should simply be left alone. Dolly in particular has become an icon, epitomising the ambivalence of biotechnology—its promise and the dilemmas it poses. The photograph of her surrounded by a flock of paparazzi at the Roslin Institute outside Edinburgh also highlights that these frontiers have been reached in an era of instant global communications. There is unprecedented media interest and thus also great public awareness of these developments. No longer can biotechnology do its work in isolation and present its results to society for an assumed approval. The near universal reaction against human cloning and the current European controversy over embryonic stem cells together highlight the fact that these are developments that call for a new type of contract between science and society. To determine the directions and priorities of such far reaching areas of the biosciences, a dialogue needs to be established with civil society. It is not sufficient to invite only the “usual partners” to the table—researchers, industrial companies, government funding agencies, patent lawyers and regulators. Increasingly, ordinary citizens are wanting to have more say in what applications should go ahead, whether there are some areas to hold back from for the time being, some to forbid altogether, and where the research itself is going. The European Commission’s recent conference “Stem Cells—Therapies for the Future” was an example, in that a two day conference had not only state of the art scientific presentations, but also substantial sessions set aside for ethical issues and for participative public discussion. This paper was presented at a similar symposium, organised at the European Molecular Biology Laboratory in Heidelberg in June 2001. It explores the ethical issues raised first by reproductive cloning and then by human stem cell research. The Society Religion and Technology Project of the Church of Scotland has been in the forefront of ethical debate on cloning and stem cells since 1996. – 8 It has been in ongoing dialogue with the Roslin scientists since 1994 and also with the wider research community, Government, European and regulatory bodies. It has also sought to promote public discussion through talks and debates, drama, and the media. The paper also draws from the work of the bioethics working group of the Conference of European Churches, of which the author is also a member. This group brings together specialists in biotechnological, medical, legal, ethical and theological fields from Protestant and Orthodox churches in several European countries, and has published two reports on these issues.