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    • 5. 发明授权
    • Plasmid-encoded neurotoxin genes in Clostridium botulinum serotype A subtypes
    • 肉毒毒素肉毒杆菌血清型A亚型中的质粒编码的神经毒素基因
    • US08435759B2
    • 2013-05-07
    • US12664800
    • 2008-06-27
    • Eric A. JohnsonKristin M. MarshallSabine PellettMarite Bradshaw
    • Eric A. JohnsonKristin M. MarshallSabine PellettMarite Bradshaw
    • C12P21/00C12N15/74
    • C12N9/52C07K14/33C12N15/74C12Y304/24069
    • The present invention provides a novel isolated plasmid, wherein the plasmid is a native plasmid found in unique C. botulinum type A strains and encode either BoNT/A3 or BoNT/A4 and BoNT/B. The present invention also provides a method of obtaining a plasmid-encoded botulinum neurotoxin and botulinum neurotoxin complex comprising the step of isolating a plasmid encoding the cntA/A or cntA/B neurotoxin gene and genes encoding protein components of the toxin complex from a C. botulinum type A strain. The inventors performed comparative analyzes of representative BoNT/A subtype strains by pulsed-field gel electrophoresis (PFGE) and Southern hybridizations with probes specific for the BoNT/A and B genes, cntA/A and cntA/B. Unexpectedly, the inventors determined that the genes encoding BoNT/A3 in the A3 strain, and BoNT/A4 and BoNT/B in the A4 strain, are on plasmids.
    • 本发明提供了一种新的分离质粒,其中质粒是在独特的A型肉毒杆菌中发现的天然质粒,并且编码BoNT / A3或BoNT / A4和BoNT / B。 本发明还提供了获得质粒编码的肉毒杆菌神经毒素和肉毒杆菌神经毒素复合物的方法,包括分离编码cntA / A或cntA / B神经毒素基因的质粒和编码来自C的毒素复合物的蛋白质组分的基因的步骤。 A型肉毒杆菌。 本发明人通过脉冲场凝胶电泳(PFGE)和针对BoNT / A和B基因cntA / A和cntA / B特异性探针的Southern杂交进行了代表性的BoNT / A亚型菌株的比较分析。 出人意料的是,本发明人确定了A3菌株中编码BoNT / A3的基因以及A4菌株中的BoNT / A4和BoNT / B在质粒上。
    • 9. 发明申请
    • Methods of Mapping Polymorphisms and Polymorphism Microarrays
    • 多态性和多态性微阵列的映射方法
    • US20080194418A1
    • 2008-08-14
    • US11914099
    • 2006-05-10
    • Eric A. JohnsonGuowen LiuMichael R. Miller
    • Eric A. JohnsonGuowen LiuMichael R. Miller
    • C40B30/04C12Q1/68C40B40/08C40B50/06
    • C12Q1/683C12Q2563/131C12Q2533/101C12Q2525/191C12Q2565/501
    • Described herein are methods for the high-throughput discovery and genotyping of nucleotide polymorphisms in DNA, including single nucleotide polymorphism (SNPs) and short deletions and insertions. These methods take advantage of the fact that differences in DNA sequence result in the differential presence of restriction endonuclease digestion sites. Differences can be detected between individuals, or the relative presence detected in a population. Provided approaches involve isolation of short DNA fragments (“tags) near restriction endonuclease sites. The presence of one (or two) of these tags indicates that a site was present. Distinguishable labeling of tags from two individual or populations allows comparative presence of these sites to be assayed on a platform that employs a collection of nucleic acids. Other approaches depend on the differential presence of restriction endonuclease sites, but involve mixing genomic DNA from the two individuals. Regions of DNA with a restriction site in only one individual create an opportunity for primer extension to produce labeled material, which can be assayed on a platform that employs a collection of nucleic acids. Any of a variety of detection platforms can be use with the described approaches. By way of example, highly efficient variant detection microarrays and bead libraries are provided that contain genomic tags with different representations between two populations, so that most elements in the collection of nucleic acids contain an informative SNP between the populations of interest.
    • 本文描述了DNA的核苷酸多态性的高通量发现和基因分型的方法,包括单核苷酸多态性(SNPs)和短缺失和插入。 这些方法利用DNA序列的差异导致限制性内切核酸酶消化位点的差异存在的事实。 可以在个体之间检测差异,或在群体中检测到相对存在。 提供的方法包括在限制性内切核酸酶位点附近分离短DNA片段(“标签”),一个(或两个)这些标签的存在表明存在一个位点。来自两个个体或群体的标签的区分标签允许这些位点的比较存在 在采用核酸集合的平台上进行检测其他方法取决于限制性内切核酸酶位点的差异存在,但涉及混合来自两个个体的基因组DNA。仅在一个个体中具有限制性位点的DNA区域产生机会 用于引物延伸以产生标记的物质,其可以在采用核酸集合的平台上进行测定。多种检测平台中的任何一种可以与所述方法一起使用。作为示例,高效变体检测微阵列和珠粒 提供了包含两个群体之间具有不同表征的基因组标签的文库,因此大多数 核酸集合中的元素包含感兴趣的群体之间的信息性SNP。