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21. 发明授权

US06403780B1 Homologous 28-kilodalton immunodominant protein genes of ehrlichia canis and uses thereof 失效
标题翻译：同源28千道尔顿免疫显性蛋白基因的埃及大肠杆菌及其用途
公开(公告)号：US06403780B1
公开(公告)日：2002-06-11
申请号：US09261358
申请日：1999-03-03
申请人： David H. Walker , Xue-Jie Yu , Jere W. McBride
发明人： David H. Walker , Xue-Jie Yu , Jere W. McBride
IPC分类号： C12N1500
CPC分类号： C07K14/29 , A61K2039/52 , Y02A50/403
摘要： The present invention is directed to the cloning, sequencing and expression of homologous immunoreactive 28-kDa protein genes, ECa28-1 and ECa28SA3, from a polymorphic multiple gene family of Ehrlichia canis. A complete sequence of another 28-kDa protein gene, ECaSA2, is also provided. Further disclosed is a multigene locus encoding all five homologous 28-kDa protein genes of Ehrlichia canis. Recombinant Ehrlichia canis 28-kDa proteins react with convalescent phase antiserum from an E. canis-infected dog.
摘要翻译：本发明涉及从埃氏杆菌的多态性多基因家族克隆，测序和表达同源免疫反应性28-kDa蛋白质基因ECa28-1和ECa28SA3。另外提供了另一个28-kDa蛋白质基因ECaSA2的完整序列。进一步公开的是编码埃里希氏菌的所有五个同源的28-kDa蛋白质基因的多基因座。重组埃氏体28-kDa蛋白质与来自犬属感染犬的恢复期抗血清反应。

22. 发明授权

US06355777B1 P43 antigen for the immunodiagnosis of canine ehrlichiosis and uses thereof 失效
标题翻译： P43抗原用于犬ehrlichiosis的免疫诊断及其用途
公开(公告)号：US06355777B1
公开(公告)日：2002-03-12
申请号：US09561322
申请日：2000-04-28
申请人： David H. Walker , Jere W. McBride
发明人： David H. Walker , Jere W. McBride
IPC分类号： C07K100
CPC分类号： C07K14/29 , A61K38/00
摘要： Canine monocytic ehrlichiosis, caused by Ehrilichia canis is a potentially fatal disease of dogs that requires rapid and accurate diagnosis in order to initiate appropriate therapy leading to a favorable prognosis. In the invention described herein, a new immunoreactive E. canis surface protein gene of 1170-bp was cloned, which encodes a protein with a predicted molecular mass of 42.6 kilodaltons (P43). The P43 gene was not found in E. chaffeensis DNA by Southern blot, and antisera against recombinant P43 (rP43) did not react with E. chaffeensis by IFA. The P43 was located on the surface of E. canis by immunoelectron microscopy. Forty-two dogs exhibiting signs and/or hematologic abnormalities associated with canine ehrlichiosis were tested by IFA and by Western immunoblot. Among the 22 samples that were IFA positive for E. canis, 100% reacted with the rP43, 96% with the rP28, and 96% with the rP140. The specificity of the recombinant proteins compared to IFA was 96% for rP28, 88% for P43 and 63% for P140. Results of this study demonstrate that the rP43 and rP28 are sensitive and reliable serodiagnostic antigens for the diagnosis of Ehrlichia canis infections.
摘要翻译：犬单核细胞性埃里希病是由艾氏杆菌引起的，是潜在致命的狗疾病，需要快速准确的诊断，以启动适当的治疗，导致有利的预后。在本文描述的本发明中，克隆了1170-bp的新的免疫反应性E.canis表面蛋白基因，其编码具有42.6千道尔顿（P43）的预测分子量的蛋白质。通过Southern印迹，在E.caffeensis DNA中没有发现P43基因，并且针对重组P43（rP43）的抗血清与IFA不与E.Chaffeensis反应。通过免疫电子显微镜观察，P43位于大肠杆菌表面。通过IFA和Western免疫印迹检测显示与犬ehrlichiosis相关的迹象和/或血液异常的42只狗。在22例IFA阳性的样本中，100％与rP43反应，96％与rP28反应，96％与rP140反应。与IFA相比，重组蛋白的特异性为rP28的96％，P43为88％，P140为63％。这项研究的结果表明，rP43和rP28是敏感和可靠的血清诊断抗原，用于诊断艾氏杆菌感染。

23. 发明授权

US4993233A Demand defrost controller for refrigerated display cases 失效
标题翻译：冷藏展示柜需求除霜控制器
公开(公告)号：US4993233A
公开(公告)日：1991-02-19
申请号：US385103
申请日：1989-07-26
申请人： David N. Borton , David H. Walker
发明人： David N. Borton , David H. Walker
IPC分类号： A47F3/04 , F25D21/00
CPC分类号： A47F3/0447 , F25D21/006 , F25B2700/21172 , F25B2700/21173
摘要： A demand defrost controller employs temperature measurements taken at the outlet of the discharge air curtain and the inlet of the air return to determine the need for defrost. If the difference between these temperature measurements exceeds a set point for a predetermined sustained period of time and a minimum amount of time has elapsed since the last defrost of the case, defrost is initiated. Regardless of the value of the temperature difference, defrost is initiated if elapsed time since the last defrost exceeds a specified maximum time between defrosts for the case. Defrost can be terminated on the basis of time and/or temperature. The demand defrost controller can also control display case temperature and can be applied to independently control multiple display case circuits.
摘要翻译：需求除霜控制器采用在排放风幕出口和空气回流入口进行的温度测量，以确定除霜的需要。如果这些温度测量值之间的差异在预定的持续时间段内超过设定点，并且自该情况的最后一次除霜以来已经过去了最少的时间量，则开始除霜。不管温度差值如何，如果从最后一次除霜以后的时间超过规定的最大时间，除霜之前就开始除霜。除霜可以根据时间和/或温度终止。需求除霜控制器还可以控制显示箱温度，并可应用于独立控制多个显示屏电路。

24. 发明申请

US20090275102A1 EHRLICHIA DISULFIDE BOND FORMATION PROTEINS AND USES THEREOF 有权
标题翻译： EHRLICHIA DISOLFIDE BOND形成蛋白及其用途
公开(公告)号：US20090275102A1
公开(公告)日：2009-11-05
申请号：US12242865
申请日：2008-09-30
申请人： David H. Walker , Jere W. McBride
发明人： David H. Walker , Jere W. McBride
IPC分类号： C12N9/02
CPC分类号： C12N9/0051 , A61K39/00 , A61K2039/522 , C07K14/29 , Y02A50/403
摘要： Novel genes encoding homologous immunoreactive thio-disulfide oxidoreductases, or disulfide bond formation (Dsb) proteins from Ehrlichia chaffeensis and Ehrlichia canis are disclosed. While the E. chaffeensis and E. canis Dsb proteins are at most only 31% or less homologous to other known Dsb proteins, the Ehrlichia Dsbs contain a cysteine active site, Cys-Gly-Tyr-Cys, similar to those in known Dsb proteins. As predicted by 15-amino acid identical N-terminal signal peptides, the proteins are primarily localized in the periplasm of E. chaffeensis and E. canis, possibly playing a role in antigenicity and pathogenesis. The present invention provides the nucleotide and amino acid sequences and expression vectors for the E. chaffeensis and E. canis dsb genes, antisera directed against the proteins, and kits to determine whether an individual or animal is infected with a given species of Ehrlichia.
摘要翻译：公开了编码同源免疫反应性硫代二硫化物氧化还原酶的新型基因或来自埃里希氏菌和埃氏杆菌的二硫键形成（Dsb）蛋白。虽然E.Chaffeensis和E. canis Dsb蛋白质与其他已知的Dsb蛋白质最多只有31％或更少的同源性，但是Ehrlichia Dsbs含有与已知Dsb蛋白质相似的半胱氨酸活性位点Cys-Gly-Tyr-Cys 。如通过15个氨基酸相同的N末端信号肽所预测的，这些蛋白质主要定位于E.Chaffeensis和E. canis的周质中，可能在抗原性和发病机理中起作用。本发明提供了E.caffeensis和E.canis dsb基因的核苷酸和氨基酸序列和表达载体，针对蛋白质的抗血清，以及用于确定个体或动物是否用给定种类的埃里希菌感染的试剂盒。

25. 发明授权

US07556816B2 Ehrlichia chaffeensis 28 kDa outer membrane protein multigene family 失效
公开(公告)号：US07556816B2
公开(公告)日：2009-07-07
申请号：US10284986
申请日：2002-10-31
申请人： David H. Walker , Xue-Jie Yu
发明人： David H. Walker , Xue-Jie Yu
IPC分类号： A61K39/02 , C07K14/295 , C07H21/04 , C12P21/04
CPC分类号： C07K14/29 , A61K39/00 , Y02A50/403
摘要： The 28-kDa outer membrane proteins (P28) of Ehrlichia chaffeensis are encoded by a multigene family consisting of 21 members located in a 23-kb DNA fragment in the genome of E. chaffeensis. Fifteen of these proteins are claimed herein as novel sequences. The amino acid sequence identity of the various P28 proteins was 20-83%. Six of 10 tested p28 genes were actively transcribed in cell culture grown E. chaffeensis. RT-PCR also indicated that each of the p28 genes was monocistronic. These results suggest that the p28 genes are active genes and encode polymorphic forms of the P28 proteins. The P28s were also divergent among different isolates of E. chaffeensis. The large repertoire of the p28 genes in a single ehrlichial organism and antigenic diversity of the P28 among the isolates of E. chaffeensis suggest that the P28s may be involved in immune avoidance.

26. 发明授权

US07432081B2 Ehrlichia disulfide bond formation proteins and uses thereof 失效
标题翻译：二硫键形成蛋白及其用途
公开(公告)号：US07432081B2
公开(公告)日：2008-10-07
申请号：US10286516
申请日：2002-11-01
申请人： David H. Walker , Jere W. McBride
发明人： David H. Walker , Jere W. McBride
IPC分类号： C12P19/34 , C12P21/04 , A61K39/02
CPC分类号： C12N9/0051 , A61K39/00 , A61K2039/522 , C07K14/29 , Y02A50/403
摘要： Novel genes encoding homologous immunoreactive thio-disulfide oxidoreductases, or disulfide bond formation (Dsb) proteins from Ehrlichia chaffeensis and Ehrlichia canis are disclosed. While the E. chaffeensis and E. canis Dsb proteins are at most only 31% or less homologous to other known Dsb proteins, the Ehrlichia Dsbs contain a cysteine active site, Cys-Gly-Tyr-Cys, similar to those in known Dsb proteins. As predicted by 15-amino acid identical N-terminal signal peptides, the proteins are primarily localized in the periplasm of E. chaffeensis and E. canis, possibly playing a role in antigenicity and pathogenesis. The present invention provides the nucleotide and amino acid sequences and expression vectors for the E. chaffeensis and E. canis dsb genes, antisera directed against the proteins, and kits to determine whether an individual or animal is infected with a given species of Ehrlichia.
摘要翻译：公开了编码同源免疫反应性硫代二硫化物氧化还原酶的新型基因或来自埃里希氏菌和埃氏杆菌的二硫键形成（Dsb）蛋白。虽然E.Chaffeensis和E. canis Dsb蛋白质与其他已知的Dsb蛋白质最多只有31％或更少的同源性，但是Ehrlichia Dsbs含有与已知Dsb蛋白质相似的半胱氨酸活性位点Cys-Gly-Tyr-Cys 。如通过15个氨基酸相同的N末端信号肽所预测的，这些蛋白质主要定位于E.Chaffeensis和E. canis的周质中，可能在抗原性和发病机理中起作用。本发明提供了E.caffeensis和E.canis dsb基因的核苷酸和氨基酸序列和表达载体，针对蛋白质的抗血清，以及用于确定个体或动物是否用给定种类的埃里希菌感染的试剂盒。

27. 发明授权

US07332171B2 Ehrlichia chaffeensis 28 kDa outer membrane protein multigene family 失效
标题翻译： Ehrlichia chaffeensis 28 kDa外膜蛋白多基因家族
公开(公告)号：US07332171B2
公开(公告)日：2008-02-19
申请号：US10369293
申请日：2003-02-18
申请人： David H. Walker , Xue-Jie Yu
发明人： David H. Walker , Xue-Jie Yu
IPC分类号： A61K39/02 , C12P21/02 , C07H21/04 , C12N15/09
CPC分类号： C07K14/29 , A61K39/00 , Y02A50/403
摘要： The 28-kDa outer membrane proteins (P28) of Ehrlichia chaffeensis are encoded by a multigene family consisting of 21 members located in a 23-kb DNA fragment in the genome of E. chaffeensis. Fifteen of these proteins are claimed herein as novel sequences. The amino acid sequence identity of the various P28 proteins was 20-83%. Six of 10 tested p28 genes were actively transcribed in cell culture grown E. chaffeensis. RT-PCR also indicated that each of the p28 genes was monocistronic. These results suggest that the p28 genes are active genes and encode polymorphic forms of the P28 proteins. The P28s were also divergent among different isolates of E. chaffeensis. The large repertoire of the p28 genes in a single ehrlichial organism and antigenic diversity of the P28 among the isolates of E. chaffeensis suggest that the P28s may be involved in immune avoidance.
摘要翻译： Ehilichia chaffeensis的28-kDa外膜蛋白（P28）由多基因家族编码，该基因由21个成员组成，位于E.caffeensis基因组的23-kb DNA片段中。这些蛋白质中的十五个被称为新的序列。各种P28蛋白的氨基酸序列同一性为20-83％。 10个测试的p28基因中有6个在生长的E.Chaffeensis的细胞培养物中被主动转录。 RT-PCR也表明每个p28基因是单顺反子。这些结果表明p28基因是活性基因，并编码P28蛋白的多态性。不同分离株之间的P28s也是不同的。大肠杆菌中单核细胞中p28基因的大部分谱系和P28的抗原多样性表明，P28可能参与免疫避免。

28. 发明授权

US06806065B2 Rickettsia felis outer membrane protein 失效
标题翻译：立克次体外膜蛋白
公开(公告)号：US06806065B2
公开(公告)日：2004-10-19
申请号：US09800065
申请日：2001-03-06
申请人： Donald H. Bouyer , Patricia Crocquet-Valdes , John Stenos , David H. Walker
发明人： Donald H. Bouyer , Patricia Crocquet-Valdes , John Stenos , David H. Walker
IPC分类号： C12N1509
CPC分类号： C07K14/29 , A61K39/00 , Y02A50/403
摘要： The present invention is directed to isolated nucleic acid molecules encoding Rickettsia felis outer membrane proteins (R. felis omp). Expression vectors and host cells comprising the nucleic acid molecules are also provided, as well as methods for increasing or decreasing the expression of R. felis omp in host cells. The invention further provides a method of screening a substance for the ability of the substance to modify R. felis omp function, and a method for isolating other R. felis omp molecules. DNA oligomers capable of hybridizing to the nucleic acid molecule encoding the R. felis omp are provided, which can be used to detect R. felis omp in a sample. An isolated R. felis omp is also provided. Antibodies specific for the protein, and fragments thereof, are provided, as are compositions comprising the protein and a compatible carrier. The subject invention further provides a method of preventing R. felis infections by R. felis present in a carrier host, and a method of reducing R. felis infection of a carrier host.
摘要翻译：本发明涉及编码立克次体外膜蛋白（R.felis omp）的分离的核酸分子。还提供了包含核酸分子的表达载体和宿主细胞，以及用于增加或降低宿主细胞中的糠the的表达的方法。本发明还提供了一种物质筛选物质修饰R.felis omp功能的方法，以及分离其他R.felis omp分子的方法。提供了能够与编码罗非鱼的核酸分子杂交的DNA寡聚体，其可以用于检测样品中的R.felis omp。还提供了一种孤立的R. felis omp。提供了对蛋白质及其片段特异性的抗体，以及包含蛋白质和相容载体的组合物。本发明进一步提供了一种防止存在于载体宿主中的R.felis的R.felis感染的方法，以及减少载体宿主感染的方法。

29. 发明授权

US06392023B1 Homologous 28-kilodalton immunodominant protein genes of Ehrlicha canis and uses thereof 失效
标题翻译： Ehrlicha canis的同源28千道尔顿免疫显性蛋白基因及其用途
公开(公告)号：US06392023B1
公开(公告)日：2002-05-21
申请号：US09660587
申请日：2000-09-12
申请人： David H. Walker , Xue-Jie Yu , Jere W. McBride
发明人： David H. Walker , Xue-Jie Yu , Jere W. McBride
IPC分类号： C12N1511
CPC分类号： C07K14/29 , A61K39/00 , A61K2039/52 , Y02A50/403
摘要： The present invention is directed to the cloning, sequencing and expression of homologous immunoreactive 28-kDa protein genes, p28-1, -2, -3, -5, -6, -7, -9, from a polymorphic multiple gene family of Ehrlichia canis. Further disclosed is a multigene locus encoding all nine homologous 28-kDa protein genes of Ehrlichia canis. Recombinant Ehrlichia canis 28-kDa proteins react with convalescent phase antiserum from an E. canis-infected dog, and may be useful in the development of vaccines and serodiagnostics that are particularly effective for disease prevention and serodiagnosis.
摘要翻译：本发明涉及来自多态性多基因家族的同源免疫反应性28-kDa蛋白基因p28-1，-2，-3，-5，-6，-7，-9的克隆，测序和表达艾氏杆菌进一步披露的是编码所有9个同源的28-kDa蛋白质基因的多基因位点。重组埃氏体28-kDa蛋白质与来自犬属感染狗的恢复期抗血清反应，并且可用于开发对预防疾病和血清诊断特别有效的疫苗和血清学诊断。

30. 发明授权

US6043085A Ehrlichia canis 120-kDa immunodominant antigenic protein and gene 有权
标题翻译：埃氏杆菌120-kDa免疫显性抗原蛋白和基因
公开(公告)号：US6043085A
公开(公告)日：2000-03-28
申请号：US141047
申请日：1998-08-27
申请人： Xue-Jie Yu , David H. Walker
发明人： Xue-Jie Yu , David H. Walker
IPC分类号： C12N15/09 , A61K38/00 , A61K39/00 , A61K39/02 , A61K48/00 , A61P31/04 , A61P37/04 , C07K14/195 , C07K14/29 , C07K16/12 , C12N1/15 , C12N1/19 , C12N1/21 , C12N5/10 , C12N15/31 , C12N15/85 , C12N15/86 , C12P21/08 , C12R1/01 , C12N15/00 , C12N1/20 , C12N1/14 , C12N1/16
CPC分类号： C07K14/29 , A61K39/00
摘要： The present invention provides a 120-kDa protein gene of Ehrlichia canis, amplified by PCR using primers derived from the DNA sequences flanking the Ehrlichia chaffeensis 120-kDa protein gene. The recombinant E. canis 120-kDa protein contains 14 tandem repeat units with 36 amino acids each. The repeat units are hydrophilic and predicted to be surface-exposed. Also disclosed is that the recombinant E. canis 120-kDa protein is antigenic and reacts with sera from dogs convalescent from canine ehrlichiosis.
摘要翻译：本发明提供了埃里氏杆菌的120-kDa蛋白质基因，通过使用衍生自侧翼于埃里希氏杆菌120-kDa蛋白质基因的DNA序列的引物通过PCR扩增。重组大肠杆菌120-kDa蛋白含有14个串联重复单元，每个具有36个氨基酸。重复单元是亲水性的并且被预测为表面暴露的。还公开的是，重组E.杆菌120-kDa蛋白质是抗原性的，并且与来自犬埃里希氏菌病康复病犬的血清反应。

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