Genetic analysis, techniques and applications最新文献

{"title":"Transgenic rabbits for the production of biologically-active recombinant proteins in the milk","authors":"Fidel Ovidio Castro,&nbsp;José Limonta,&nbsp;Alina Rodrı́guez,&nbsp;Alina Aguirre,&nbsp;José de la Fuente,&nbsp;Anselmo Aguilar,&nbsp;Boris Ramos,&nbsp;Orlando Hayes","doi":"10.1016/S1050-3862(99)00024-8","DOIUrl":"10.1016/S1050-3862(99)00024-8","url":null,"abstract":"<div><p>The use of live bioreactors for the expression of human genes in the mammary gland of transgenic animals is one of the most cost-effective ways for the production of valuable recombinant therapeutic proteins. Among the transgenic species used so far, rabbits are good candidates for the expression of tens to hundreds of grams of complex proteins in the milk during lactation. The lactating mammary gland of rabbits has proven to be effective in the processing of complex proteins. In this work, the potential use of rabbits as bioreactors is discussed based on our results and the published data.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 3","pages":"Pages 179-187"},"PeriodicalIF":0.0,"publicationDate":"1999-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(99)00024-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21454370","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Immunological control of ectoparasites: past achievements and future research priorities","authors":"Peter Willadsen","doi":"10.1016/S1050-3862(99)00016-9","DOIUrl":"10.1016/S1050-3862(99)00016-9","url":null,"abstract":"<div><p>Recombinant vaccines are available for the control of the tick <em>Boophilus microplus</em>, while progress has been made in the development of vaccines against <em>Lucilia cuprina</em> and <em>Chrysomya bezziana</em>. Literature suggests that the control of other ectoparasites is feasible, either through the duplication in a vaccine of naturally acquired immunity or through ‘concealed’ antigen vaccines. Major deficiencies in our current knowledge however point to possible research opportunities for the future. The identification of protective antigens from all species is proceeding slowly, particularly for the antigens of naturally acquired immunity. Our capacity to produce effective recombinant antigens has progressed greatly, though there remains a major difficulty where some or all of the protective effect is due to immunogenic oligosaccharide. Our understanding of protective mechanisms is limited. The delivery of the appropriate immunological response remains difficult. Nevertheless, some of the most critical areas of ignorance are in basic biological issues: factors which affect the susceptibility of particular pest species to immunological attack and the implications of vaccine-induced effects for pest and disease control under field conditions. Increasingly too, effective pest control is likely to demand the integration of a variety of control technologies. The study of this integration is in its infancy.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 3","pages":"Pages 131-137"},"PeriodicalIF":0.0,"publicationDate":"1999-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(99)00016-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21454425","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evidence and mechanisms of immunosuppression in tick infestations","authors":"Omar O Barriga","doi":"10.1016/S1050-3862(99)00017-0","DOIUrl":"10.1016/S1050-3862(99)00017-0","url":null,"abstract":"<div><p>The abundance and ubiquity of ticks from ancient times long ago suggested that they have eluded host immunity. In the last 15 years, several authors have demonstrated suppression of the Th1 responses (cell-mediated immunity), and sometimes the Th2 responses (humoral immunity), subsequent to tick infestations in laboratory and natural models. Although the mechanisms to produce suppression are not well-defined yet, evidences for antigenic competition, lymphocyte cytotoxicity, presence of immuno-inhibiting substances in the saliva, and existence of modulators of cytokines in salivary extracts have been reported. Management of tick-induced immunosuppression is essential to replace tick control by acaricide application with more environmentally sound vaccination.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 3","pages":"Pages 139-142"},"PeriodicalIF":0.0,"publicationDate":"1999-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(99)00017-0","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21454426","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The use of yeast artificial chromosomes in transgenic animals: expression studies of the tyrosinase gene in transgenic mice","authors":"Patricia Giraldo,&nbsp;Estela Giménez,&nbsp;Lluı́s Montoliu","doi":"10.1016/S1050-3862(99)00023-6","DOIUrl":"10.1016/S1050-3862(99)00023-6","url":null,"abstract":"<div><p>Variegation and inherited somatic mosaicism has been observed in transgenic mice carrying yeast artificial chromosomes (YACs) in which a DNAse I hypersensitive site (HS) located −12 kb upstream of the mouse tyrosinase gene had been deleted. At present, we are generating new transgenic animals with minor deletions of the HS.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 3","pages":"Pages 175-178"},"PeriodicalIF":0.0,"publicationDate":"1999-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(99)00023-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21454431","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Transgenic pigs as bioreactors: a comparison of gamma-carboxylation of glutamic acid in recombinant human protein C and factor IX by the mammary gland","authors":"Kevin E Van Cott ,&nbsp;Stephen P Butler ,&nbsp;Christopher G Russell ,&nbsp;Anu Subramanian ,&nbsp;Henryk Lubon ,&nbsp;F.C Gwazdauskas ,&nbsp;James Knight ,&nbsp;William N Drohan ,&nbsp;William H Velander","doi":"10.1016/S1050-3862(99)00020-0","DOIUrl":"10.1016/S1050-3862(99)00020-0","url":null,"abstract":"<div><p>The mammary gland of transgenic livestock can be used as a bioreactor for producing complex therapeutic proteins. However, the capacity for making a given post-translational modification upon any given polypeptide is uncertain. For example, the efficiency of gamma-carboxylation of glutamic acid in the amino terminal regions of recombinant human protein C (rhPC) and recombinant human Factor IX (rhFIX) is different at similar expression levels. At an expression level of about 200 μg/ml in the milk of transgenic pigs, rhFIX is highly gamma-carboxylated as indicated by pro-coagulant activity and amino acid sequencing. However, only about 20–35% of rhPC has a native, gamma-carboxyglutamic acid-dependent conformation and <em>anti</em>-coagulant activity. Thus, this work provides an example of apparent differences in substrate specificity between two homologous proteins to the endogenous carboxylase of porcine mammary epithelium which leads to varying degrees of post-translational modification.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 3","pages":"Pages 155-160"},"PeriodicalIF":0.0,"publicationDate":"1999-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(99)00020-0","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21454430","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biotecnologia Habana ‘98: Latin America in the Collimator","authors":"Ana Cristina Campal Espinosa","doi":"10.1016/S1050-3862(99)00009-1","DOIUrl":"10.1016/S1050-3862(99)00009-1","url":null,"abstract":"","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 3","pages":"Pages 73-74"},"PeriodicalIF":0.0,"publicationDate":"1999-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(99)00009-1","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21454520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The optimal use of IRES (internal ribosome entry site) in expression vectors","authors":"Joe Attal,&nbsp;Marie-Claire Théron,&nbsp;Louis Marie Houdebine","doi":"10.1016/S1050-3862(99)00021-2","DOIUrl":"10.1016/S1050-3862(99)00021-2","url":null,"abstract":"<div><p>In higher eucaryotes, natural bicistronic mRNA have been rarely found so far. The second cistron of constructed bicistronic mRNAs is generally considered as not translated unless special sequences named internal ribosome entry site (IRES) are added between the two cistrons. These sequences are believed to recruit ribosomes independently of a cap structure. In the present report, a new IRES found in the HTLV-1 genome is described. A systematic study revealed that this IRES, but also the poliovirus (polio) and the encephalomyocarditis virus (EMCV) IRES work optimally when they are added about 100 nucleotides after the termination codon of the first cistron. Unexpectedly, these IRES became totally inefficient when added after 300–500 nucleotide spacers. This result and others are not compatible with the admitted mechanism of IRES action. The IRES appear to be rather potent translation stimulators. Their effects are particularly emphasized in cells in which the normal mechanism of translation initiation is inhibited. For these reasons, we suggest to call IRES rescue translation stimulators (RTS).</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 3","pages":"Pages 161-165"},"PeriodicalIF":0.0,"publicationDate":"1999-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(99)00021-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21454429","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Vaccination against ticks (Boophilus spp.): the experience with the Bm86-based vaccine Gavac™","authors":"José de la Fuente ,&nbsp;Manuel Rodrı́guez ,&nbsp;Carlos Montero ,&nbsp;Miguel Redondo ,&nbsp;José Carlos Garcı́a-Garcı́a ,&nbsp;Luis Méndez ,&nbsp;Emerio Serrano ,&nbsp;Mario Valdés ,&nbsp;Antonio Enrı́quez ,&nbsp;Mario Canales ,&nbsp;Eduardo Ramos ,&nbsp;Oscar Boué ,&nbsp;Héctor Machado ,&nbsp;Ricardo Lleonart","doi":"10.1016/S1050-3862(99)00018-2","DOIUrl":"10.1016/S1050-3862(99)00018-2","url":null,"abstract":"<div><p>The control of tick infestations and the transmission of tick-borne diseases remain a challenge for the cattle industry in tropical and subtropical areas of the world. Traditional control methods have been only partially successful and the parasites continue to result in significant losses for the cattle industry. Recently, vaccines containing the recombinant <em>B. microplus</em> gut antigen Bm86 have been developed. Our vaccine formulation (Gavac™, Heber Biotec S.A., Havana, Cuba) has been registered and is commercially available in Cuba, Colombia, Dominican Republic, Brazil and Mexico. In controlled pen trials, Gavac™ has been effective for the control of artificial infestations of <em>B. annulatus</em>, <em>B. decoloratus</em> and chemical-sensitive and resistant <em>B. microplus</em> strains from Australia, Africa, America and Iran. In controlled field trials in Cuba, Brazil, Argentina and Mexico, Gavac has shown a 55–100% efficacy in the control of <em>B. microplus</em> infestations in grazing cattle 12–36 weeks after the first vaccination. Field trials under production conditions have been conducted in Cuba, Colombia, Brazil and Mexico in pure and cross-bred cattle herds. The application of Gavac™ has increased the time between acaricide treatments by an average of 32±21 days (<em>P</em>=0.0005) resulting in important savings for the cattle industry. In Cuba, a cost-effectiveness analysis was conducted in more than 260 000 animals. The cost-effectiveness analysis showed a 60% reduction in the number of acaricide treatments, together with the control of tick infestations and transmission of babesiosis, which resulted in savings of $23.4 animal⁻¹ year⁻¹. These results clearly demonstrate the advantage of vaccination and support the application of Gavac for the control of <em>Boophilus</em> spp. infestations.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 3","pages":"Pages 143-148"},"PeriodicalIF":0.0,"publicationDate":"1999-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(99)00018-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21454427","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PCR based targeted genomic and cDNA differential display","authors":"NataliaE. Broude ,&nbsp;Niels Storm ,&nbsp;Sarah Malpel ,&nbsp;JoelH. Graber ,&nbsp;Sergey Lukyanov ,&nbsp;Eugene Sverdlov ,&nbsp;CassandraL. Smith","doi":"10.1016/S1050-3862(98)00038-2","DOIUrl":"10.1016/S1050-3862(98)00038-2","url":null,"abstract":"<div><p>We previously described a targeted genomic differential display method (TGDD: Broude NE, Chandra A, Smith CL. Differential display of genomic subsets containing specific interspersed repeats. Proc. Natl. Acad. Sci. USA 1997;94:4548–53). In that method, presently characterized as method I, targeting was accomplished by capturing DNA fragments containing specific a sequence by hybridization with complementary single-stranded DNA. The captured fragments were amplified by PCR. Here, we describe method II where targeting is accomplished by PCR using primers specific to the target sequence. Method II takes advantage of PCR suppression to eliminate fragments not containing the target sequence (Siebert PDA, Chenchik A, Kellogg DE, Lukyanov KA and Lukyanov SA. An improved PCR method for walking in uncloned genomic DNA. Nucleic Acids Res 1995;23:1087–1088). Targeting focuses analysis on and around interesting areas and additionally serves to reduce the complexity of the amplified subset. These approaches are useful to amplify genome subsets containing a variety of targets including various conserved sequences coding for <em>cis</em>-acting elements or protein motifs.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 2","pages":"Pages 51-63"},"PeriodicalIF":0.0,"publicationDate":"1999-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(98)00038-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21063710","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selection analysis of single-stranded clones using solid-phase techniques","authors":"Tatiana Vener,&nbsp;Anders Holmberg,&nbsp;Joakim Lundeberg,&nbsp;Mathias Uhlén","doi":"10.1016/S1050-3862(98)00035-7","DOIUrl":"10.1016/S1050-3862(98)00035-7","url":null,"abstract":"<div><p>An efficient selection analysis of single-stranded DNA fragments using subtractive hybridization on magnetic beads and fluorescence-based fragment analysis is described. This approach facilitates either direct cloning of captured fragments or removal of common clones from cDNA or shot-gun clone libraries.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"15 2","pages":"Pages 47-50"},"PeriodicalIF":0.0,"publicationDate":"1999-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(98)00035-7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"21063812","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}