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    Construction of recombinant DNA by exonuclease recession
    1.
    发明授权
    Construction of recombinant DNA by exonuclease recession 失效
    通过外切核酸酶衰退构建重组DNA

    公开(公告)号：US5580759A

    公开(公告)日：1996-12-03

    申请号：US192300

    申请日：1994-02-03

    申请人： Yih-Sheng Yang ,  Philip W. Tucker ,  J. Donald Capra

    发明人： Yih-Sheng Yang ,  Philip W. Tucker ,  J. Donald Capra

    IPC分类号： C12N15/10 ,  C12Q1/68 ,  C07H21/04 ,  C12P19/34

    CPC分类号： C12N15/10 ,  C12Q1/686

    摘要： An exonuclease-based method for joining and/or constructing two or more DNA molecules. DNA fragments containing ends complementary to those of a vector or another independent molecule were generated by the polymerase chain reaction. The 3' ends of these molecules as well as the vector DNA were then recessed by exonuclease activity and annealed in an orientation-determined manner via their complementary single-stranded regions. This recombinant DNA may be transformed directly into bacteria without a further ligase-dependent reaction. Using this approach, recombinant DNA molecules are constructed rapidly, efficiently and directionally. This method can effectively replace conventional protocols for PCR cloning, PCR SOEing, DNA subcloning and site-directed mutagenesis.

    摘要翻译： 用于连接和/或构建两个或多个DNA分子的基于外切核酸酶的方法。 通过聚合酶链反应产生含有与载体或另一独立分子的末端互补的末端的DNA片段。 然后将这些分子的3'末端以及载体DNA通过核酸外切酶活性凹入，并通过它们的互补单链区域以取向决定的方式进行退火。 这种重组DNA可以直接转化成细菌而不需要进一步的连接酶依赖性反应。 使用这种方法，快速，有效和定向地构建重组DNA分子。 该方法可以有效地代替PCR克隆，PCR SOEing，DNA亚克隆和定点突变的常规方案。

    Rapid generation of antibodies
    2.
    发明授权
    Rapid generation of antibodies 有权
    快速产生抗体

    公开(公告)号：US08563305B2

    公开(公告)日：2013-10-22

    申请号：US12433832

    申请日：2009-04-30

    申请人： Rafi Ahmed ,  Joseph Miller ,  Patrick C. Wilson ,  J. Donald Capra ,  Jens Wrammert

    发明人： Rafi Ahmed ,  Joseph Miller ,  Patrick C. Wilson ,  J. Donald Capra ,  Jens Wrammert

    IPC分类号： C12N5/06 ,  C12N5/02

    CPC分类号： C07K16/1018 ,  C07K16/00

    摘要： High efficient methods for producing an antibody molecule that binds an antigen are described. The methods include obtaining a population of PBMC enriched for CD19highCD3negCD20low to negCD38highCD27high cells from a mammal exposed to an antigen from sample of cells enriched for PBMC. The cells are isolated from a sample obtained at a time that the fraction of PBMC expressing antibody reactive to the antigen is at a high level. Sequences encoding heavy and light chain variable domains are prepared in a manner that allow production of molecules with natural heavy and light chain pairing.

    摘要翻译： 描述了用于产生结合抗原的抗体分子的高效方法。 所述方法包括从富集PBMC的细胞样品中获得富含CD19highCD3negCD20的PBMC群体，从暴露于抗原的哺乳动物中将CD38highCD27高细胞从抗原中除去。 从在与抗原反应的表达抗体的PBMC的分数处于高水平的时刻获得的样品中分离细胞。 编码重链和轻链可变结构域的序列以允许生成具有天然重链和轻链配对的分子的方式制备。

    DNA segments encoding autoimmune polypeptide epitopes of Ro/ss-A antigen
    3.
    发明授权
    DNA segments encoding autoimmune polypeptide epitopes of Ro/ss-A antigen 失效
    编码Ro /β-A抗原自身免疫多肽表位的DNA段

    公开(公告)号：US06303339B1

    公开(公告)日：2001-10-16

    申请号：US07576423

    申请日：1991-05-24

    申请人： Richard D. Sontheimer ,  Tsu-San Lieu ,  Daniel P. McCauliffe ,  J. Donald Capra

    发明人： Richard D. Sontheimer ,  Tsu-San Lieu ,  Daniel P. McCauliffe ,  J. Donald Capra

    IPC分类号： C12P2104

    CPC分类号： G01N33/564 ,  C07K14/46 ,  C12Q1/6876 ,  G01N2800/24

    摘要： The present disclosure relates to DNA sequences encoding one or more antigenic epitopes of the Ro 60 kD autoantigen, as well as to antigenic peptides themselves which correspond antigenically to epitopes found on the Ro/SS-A ribonucleoprotein (RNP) particle. Peptides which incorporate the antigenic epitopic core sequences disclosed herein may be employed in place of the Ro/SS-A RNP in any of a variety immunoassays including ELISA assays. The polypeptides of the invention may be employed in colorimetric assays for the identification and characterization of autoimmune diseases such as systemic lupus erythematosus (SLE) and Sjogren's syndrome. The DNA sequences disclosed herein may be employed in the preparation of the 60 kD Ro antigen, peptides which incorporate antigenic core sequences thereof, to probe for Ro sequences by hybridization analysis, and the like.

    摘要翻译： 本公开内容涉及编码Ro 60kD自身抗原的一种或多种抗原表位的DNA序列，以及抗原肽本身对应于在Ro / SS-A核糖核蛋白（RNP）颗粒上发现的表位的抗原性肽。 可以使用包含本文公开的抗原性表位核心序列的肽来代替Ro / SS-A RNP，其包括ELISA测定中的任一种免疫测定。 本发明的多肽可用于鉴定和表征自身免疫疾病如系统性红斑狼疮（SLE）和干燥综合征的比色测定。 本文公开的DNA序列可以用于制备60kD Ro抗原，其中包含其抗原核心序列的肽，通过杂交分析来探测Ro序列等。