公开(公告)号：US07449310B2

公开(公告)日：2008-11-11

申请号：US10647057

申请日：2003-08-22

Applicant: Tiruvoor G. Nagaraja ,  George C. Stewart ,  Sanjeev K. Narayanan ,  Muckatira M. Chengappa

Inventor： Tiruvoor G. Nagaraja ,  George C. Stewart ,  Sanjeev K. Narayanan ,  Muckatira M. Chengappa

IPC: C12P21/04 ,  C12P21/06 ,  C07H21/04 ,  C12N1/20 ,  A61K39/02

CPC classification number: C07K14/195 ,  A61K39/00

Abstract: The F. necrophorum gene expressing leukotoxin was sequenced and cloned. The leukotoxin open reading frame (lktA) is part of a multi-gene operon containing 9,726 bp, and encoding a protein containing 3,241 amino acids with an overall molecular weight of 335,956 daltons. The protein encoded by the gene was truncated into five polypeptides having overlapping regions by truncating the full length gene into five different sections and amplifying, expressing, and recovering the protein encoded by each of these sections. Additionally, a region upstream of the gene was sequenced and the polypeptide encoded by that nucleotide sequence was purified and isolated. These polypeptides along with the full length protein are then tested to determine their immunogenicity and protective immunity in comparison to the efficacy of immunization conferred by inactivated native leukotoxin in F. necrophorum culture supernatant.

Abstract translation: 对表达白细胞毒素的死亡细胞基因组进行测序并克隆。 白细胞介素阅读框（lktA）是含有9,726bp的多基因操纵子的一部分，编码总共分子量为335,956道尔顿的3,241个氨基酸的蛋白质。 将该基因编码的蛋白质通过将全长基因截断成5个不同部分并扩增，表达和回收由这些部分中的每一个编码的蛋白质，将该基因编码的蛋白质截短为5个具有重叠区域的多肽。 另外，对基因上游的区域进行测序，并且纯化并分离由该核苷酸序列编码的多肽。 然后测试这些多肽与全长蛋白质相比，以确定它们的免疫原性和保护性免疫，与灭活的天然白细胞介素在培养物上清液中赋予的免疫功效相比。

公开(公告)号：US20090117142A1

公开(公告)日：2009-05-07

申请号：US12171985

申请日：2008-07-11

Applicant: Tiruvoor G. Nagaraja ,  George C. Stewart ,  Sanjeev K. Narayanan ,  Muckatira M. Chengappa

Inventor： Tiruvoor G. Nagaraja ,  George C. Stewart ,  Sanjeev K. Narayanan ,  Muckatira M. Chengappa

IPC: A61K39/114 ,  C12N15/11 ,  C12N15/00

CPC classification number: C07K14/195 ,  A61K39/00

Abstract: The F. necrophorum gene expressing leukotoxin was sequenced and cloned. The leukotoxin open reading frame (lktA) is part of a multi-gene operon containing 9,726 bp, and encoding a protein containing 3,241 amino acids with an overall molecular weight of 335,956 daltons. The protein encoded by the gene was truncated into five polypeptides having overlapping regions by truncating the full length gene into five different sections and amplifying, expressing, and recovering the protein encoded by each of these sections. Additionally, a region upstream of the gene was sequenced and the polypeptide encoded by that nucleotide sequence was purified and isolated. These polypeptides along with the full length protein are then tested to determine their immunogenicity and protective immunity in comparison to the efficacy of immunization conferred by inactivated native leukotoxin in F. necrophorum culture supernatant.

Abstract translation: 对表达白细胞毒素的死亡细胞基因组进行测序并克隆。 白细胞介素阅读框（lktA）是含有9,726bp的多基因操纵子的一部分，编码总共分子量为335,956道尔顿的3,241个氨基酸的蛋白质。 将该基因编码的蛋白质通过将全长基因截断成5个不同部分并扩增，表达和回收由这些部分中的每一个编码的蛋白质，将该基因编码的蛋白质截短为5个具有重叠区域的多肽。 另外，对基因上游的区域进行测序，并且纯化并分离由该核苷酸序列编码的多肽。 然后测试这些多肽与全长蛋白质相比，以确定它们的免疫原性和保护性免疫，与灭活的天然白细胞介素在培养物上清液中赋予的免疫功效相比。

公开(公告)号：US5455034A

公开(公告)日：1995-10-03

申请号：US78066

申请日：1993-06-18

Applicant: Tiruvoor G. Nagaraja ,  Muckatira M. Chengappa

Inventor： Tiruvoor G. Nagaraja ,  Muckatira M. Chengappa

IPC: A61K39/00 ,  C07K14/195 ,  A61K39/02

CPC classification number: C07K14/195 ,  A61K39/00 ,  Y10S435/822

Abstract: A method is provided for the enhanced elaboration of leukotoxin from F. necrophorum, and subsequent production of an inactivated leukotoxoid ruminant animal vaccine against F. necrophorum infection and consequent liver abscesses and/or foot rot in such animals. The method involves forming a culture of F. necrophorum bacteria in growth media, allowing the bacteria to grow therein and to simultaneously elaborate leukotoxin in a supernate; the culturing is preferably carried out at a temperature of from about 35.degree.-41.degree. C., a pH of from about 6.5-8, and for a period of from about 4-9 hours. At the end oil of the culturing, bacterial growth and leukotoxin elaboration are terminated, preferably by separating the leukotoxin supernate, whereupon the vaccine is produced by inactivation of at least the supernate.

Abstract translation: 提供了一种用于增强拟真球菌的白细胞毒素的方法，随后生产灭活的白血病类反刍动物疫苗，用于抵抗F.necrophorum感染和随后的这些动物的肝脏脓肿和/或脚腐烂。 该方法包括在生长培养基中形成坏死性细菌培养物，使细菌在其中生长并同时在上清液中制备白细胞毒素; 培养优选在约35〜-4℃的温度，约6.5-8的pH和约4-9小时的时间内进行。 最后，终止培养，细菌生长和白细胞毒素制备的油，优选通过分离白细胞上清液，从而通过使至少上清液失活而产生疫苗。

公开(公告)号：US06669940B2

公开(公告)日：2003-12-30

申请号：US09841786

申请日：2001-04-24

Applicant: Tiruvoor G. Nagaraja ,  George C. Stewart ,  Sanjeev K. Narayanan ,  Muckatira M. Chengappa

Inventor： Tiruvoor G. Nagaraja ,  George C. Stewart ,  Sanjeev K. Narayanan ,  Muckatira M. Chengappa

IPC: A61K3902

CPC classification number: C07K14/195 ,  A61K39/00

Abstract: The F. necrophorum gene expressing leukotoxin was sequenced and cloned. The leukotoxin open reading frame (lktA) is part of a multi-gene operon containing 9,726 bp, and encoding a protein containing 3,241 amino acids with an overall molecular weight of 335,956 daltons. The protein encoded by the gene was truncated into five polypeptides having overlapping regions by truncating the full length gene into five different sections and amplifying, expressing, and recovering the protein encoded by each of these sections. Additionally, a region upstream of the gene was sequenced and the polypeptide encoded by that nucleotide sequence was purified and isolated. These polypeptides along with the full length protein are then tested to determine their immunogenicity and protective immunity in comparison to the efficacy of immunization conferred by inactivated native leukotoxin in F. necrophorum culture supernatant.

公开(公告)号：US5861162A

公开(公告)日：1999-01-19

申请号：US483382

申请日：1995-06-07

Applicant: Tiruvoor G. Nagaraja ,  Muckatira M. Chengappa

Inventor： Tiruvoor G. Nagaraja ,  Muckatira M. Chengappa

IPC: A61K39/05 ,  A61K39/114 ,  A61K39/116 ,  A61K39/02

CPC classification number: A61K39/114 ,  A61K39/05 ,  A61K2039/521 ,  A61K2039/552 ,  A61K2039/70

Abstract: Novel inocula for administration to ruminant animals such as cattle or sheep are provided in order to immunize the animals and lessen the incidence of liver abscesses and/or foot rot therein. In one aspect, the invention pertains to an A. pyogenes-derived vaccine including an inactivated cell culture product (e.g., cell-elaborated supernatant) from A. pyogenes cell culture in a suitable carrier. In another aspect, the invention relates to a multivalent vaccine including at least first and second bacterial components in a carrier; the first component comprises an inactivated cell culture product of A. pyogenes whereas the second component comprises an inactivated cell culture product of F. necrophorum. The inocula of the present invention find particular utility in incidences where ruminant animals are particularly subject to A. pyogenes infection leading to liver abscesses and/or foot rot, e.g., where the animals are regularly treated with an antibiotic or where cattle are fed a high grain content concentrate diet.

Abstract translation: 提供用于给予反刍动物例如牛或绵羊的新型接种物，以免疫动物并减轻其中肝脏脓肿和/或脚腐烂的发生率。 一方面，本发明涉及一种源自化脓性链球菌来源的疫苗，其包括在合适的载体中来自A.化脓杆菌细胞培养物的灭活的细胞培养产物（例如细胞培养的上清液）。 另一方面，本发明涉及在载体中至少包含第一和第二细菌成分的多价疫苗; 第一组分包括A.化脓性链球菌的灭活细胞培养产物，而第二组分包括灭活细菌的灭活的细胞培养产物。 本发明的接种物特别适用于反刍动物特别受A.导致肝脓肿和/或脚腐的化脓性感染的作用，例如，动物经常用抗生素治疗或牛喂养高 粮食浓缩饮食。

公开(公告)号：US5492694A

公开(公告)日：1996-02-20

申请号：US333767

申请日：1994-11-03

Applicant: Tiruvoor G. Nagaraja ,  Muckatira M. Chengappa

Inventor： Tiruvoor G. Nagaraja ,  Muckatira M. Chengappa

IPC: A61K39/00 ,  C07K14/195 ,  C12P21/04 ,  A61K39/02

CPC classification number: C07K14/195 ,  A61K39/00 ,  Y10S435/822

Abstract: A method is provided for the enhanced elaboration of leukotoxin from F. necrophorum, and subsequent production of an inactivated leukotoxoid ruminant animal vaccine against F. necrophorum infection and consequent liver abscesses and/or foot rot in such animals. The method involves forming a culture of F. necrophorum bacteria in growth media, allowing the bacteria to grow therein and to simultaneously elaborate leukotoxin in a supernate; the culturing is preferably carried out at a temperature of from about 35.degree.-41.degree. C., a pH of from about 6.5-8, and for a period of from about 4-9 hours. At the end of the culturing, bacterial growth and leukotoxin elaboration are terminated, preferably by separating the leukotoxin supernate, whereupon the vaccine is produced by inactivation of at least the supernate.

Abstract translation: 提供了一种用于增强拟真球菌的白细胞毒素的方法，随后生产灭活的白血病类反刍动物疫苗，用于抵抗F.necrophorum感染和随后的这些动物的肝脏脓肿和/或脚腐烂。 该方法包括在生长培养基中形成坏死性细菌培养物，使细菌在其中生长并同时在上清液中制备白细胞毒素; 培养优选在约35〜-4℃的温度，约6.5-8的pH和约4-9小时的时间内进行。 在培养结束时，终止细菌生长和白细胞毒素的制备，优选通过分离白细胞上清液，从而通过使至少上清液失活而产生疫苗。