An extended 5-generation family has been investigated in which 32 of the 111 family members were diagnosed as having retinitis pigmentosa (RP) The proband was a 58-year old male in whom night-blindness was first obs...An extended 5-generation family has been investigated in which 32 of the 111 family members were diagnosed as having retinitis pigmentosa (RP) The proband was a 58-year old male in whom night-blindness was first observed in early childhood, with almost loss of vision by 52 years of age The symptoms observed in other family members included night-blindness, impaired vision and visual field loss Dementia, digital abnormalities, deaf-mutism and mental retardation were variously diagnosed in a number of individuals with RP The affected and unaffected family members were tested for mutations in a range of candidate genes The 8 exons of three candidate genes have been analyzed by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing techniques A novel mutation was identified in the rhodopsin gene at codon 52 of exon 1 (TTC-TAC) that resulted in a substitution of Phe to Tyr展开更多
Next-generation sequencing （NGS） technologies allow the cost-effective sequencing of whole genomes and have expanded the scope of genomics to novel applications, such as the genome-wide characterization of intraspec...Next-generation sequencing （NGS） technologies allow the cost-effective sequencing of whole genomes and have expanded the scope of genomics to novel applications, such as the genome-wide characterization of intraspecific polymorphisms and the rapid mapping and identification of point mutations. Next-generation sequencing platforms, such as the Illumina HiSeq2ooo platform, are now commercially available at affordable prices and routinely produce an enormous amount of sequence data, but their wide use is often hindered by a lack of knowledge on how to manipulateand process the information produced. In this review, we focus on the strategies that are available to geneticists who wish to incorporate these novel approaches into their research but who are not familiar with the necessary bioinformatic concepts and computational tools. In particular, we comprehensively summarize case studies where the use of NGS technologies has led to the identification of point mutations, a strategy that has been dubbed ＂mapping-by-sequencing＇, and review examples from plants and other model species such as Caenorhabditis elegans, Saccharomyces cerevisiae, and Drosophila mela- nogaster. As these technologies are becoming cheaper and more powerful, their use is also expanding to allow mutation identification in species with larger genomes, such as many crop plants.展开更多
文摘An extended 5-generation family has been investigated in which 32 of the 111 family members were diagnosed as having retinitis pigmentosa (RP) The proband was a 58-year old male in whom night-blindness was first observed in early childhood, with almost loss of vision by 52 years of age The symptoms observed in other family members included night-blindness, impaired vision and visual field loss Dementia, digital abnormalities, deaf-mutism and mental retardation were variously diagnosed in a number of individuals with RP The affected and unaffected family members were tested for mutations in a range of candidate genes The 8 exons of three candidate genes have been analyzed by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and DNA sequencing techniques A novel mutation was identified in the rhodopsin gene at codon 52 of exon 1 (TTC-TAC) that resulted in a substitution of Phe to Tyr
基金supported by grants from the Ministerio de Economiay Competitividad of Spain(BFU2011-22825 and CSD2007-00057(TRANSPLANTA))the Generalitat Valenciana(PROMETEOII/2014/003)+2 种基金H.C.was a recipient of a Marie Curie International Reintegration Grant(PIRG03-GA-2008-231073)Research in the laboratory of H.C.was supported by a grant from the Ministerio de Economiay Competitividad of Spain(BFU2012-31719)R.C.S.held a fellowship from the Ministerio de Economfa y Competitividad of Spain(BES-2009-014106)
文摘Next-generation sequencing （NGS） technologies allow the cost-effective sequencing of whole genomes and have expanded the scope of genomics to novel applications, such as the genome-wide characterization of intraspecific polymorphisms and the rapid mapping and identification of point mutations. Next-generation sequencing platforms, such as the Illumina HiSeq2ooo platform, are now commercially available at affordable prices and routinely produce an enormous amount of sequence data, but their wide use is often hindered by a lack of knowledge on how to manipulateand process the information produced. In this review, we focus on the strategies that are available to geneticists who wish to incorporate these novel approaches into their research but who are not familiar with the necessary bioinformatic concepts and computational tools. In particular, we comprehensively summarize case studies where the use of NGS technologies has led to the identification of point mutations, a strategy that has been dubbed ＂mapping-by-sequencing＇, and review examples from plants and other model species such as Caenorhabditis elegans, Saccharomyces cerevisiae, and Drosophila mela- nogaster. As these technologies are becoming cheaper and more powerful, their use is also expanding to allow mutation identification in species with larger genomes, such as many crop plants.
