公开(公告)号：US20050186564A1

公开(公告)日：2005-08-25

申请号：US11099302

申请日：2005-04-05

Applicant: Gunter Stempfer ,  Jorg Windisch ,  Franz Knauseder

Inventor： Gunter Stempfer ,  Jorg Windisch ,  Franz Knauseder

IPC: C12P21/02 ,  C12N1/21 ,  C12N9/50 ,  C12N15/57 ,  C12N15/62 ,  C12P21/06 ,  C12R1/19 ,  C12Q1/70 ,  C07H21/04 ,  C12N9/12 ,  C12N15/74

CPC classification number: C12N15/62 ,  C07K2319/00 ,  C07K2319/21 ,  C07K2319/50 ,  C07K2319/75 ,  C12N9/506 ,  C12P21/06

Abstract: The present invention relates to a process for the recombinant production of a desired heterologous polypeptide with a clearly defined homogenous N-terminus in a bacterial host cell. A fusion protein comprising a peptide with the autoproteolytic activity of an autoprotease Npro of a pestivirus and the heterologous polypeptide is initially expressed in the form of cytoplasmic inclusion bodies in the host cell, the inclusion bodies are isolated and subsequently treated so that the desired heterologous polypeptide is cleaved autoproteolytically by the Npro activity of the fusion protein.

Abstract translation: 本发明涉及在细菌宿主细胞中重组产生具有明确界定的均质N末端的所需异源多肽的方法。 包含蚜虫病毒和异源多肽的自身蛋白酶Nspo的自身蛋白水解活性的肽的融合蛋白最初以宿主细胞中的细胞质包涵体的形式表达，分离包涵体， 随后进行处理，使得所需的异源多肽被融合蛋白的N-proPAT活性自发蛋白酶切割。

公开(公告)号：US07378512B2

公开(公告)日：2008-05-27

申请号：US10763619

申请日：2004-01-23

Applicant: Tillmann Rumenapf ,  Heinz J Thiel ,  Jorg Windisch ,  Franz Knauseder

Inventor： Tillmann Rumenapf ,  Heinz J Thiel ,  Jorg Windisch ,  Franz Knauseder

IPC: C12N15/62 ,  C12N15/57 ,  C12N15/40 ,  C12N15/70 ,  C12N9/48

CPC classification number: C12P21/06 ,  C07K2319/00 ,  C07K2319/50 ,  C07K2319/75 ,  C12N9/506 ,  C12N15/62

Abstract: The present invention relates to a process for the production of a heterologous polypeptide with homogeneous N-terminus in a bacterial host cell, wherein the heterologous polypeptide is autoproteolytically cleaved from an expressed fusion protein which comprises a polypeptide with the autoproteolytic activity of an autoprotease Npro of a pestivirus and the heterologous polypeptide by the Npro autoproteolytic activity.

Abstract translation: 本发明涉及用于在细菌宿主细胞中产生具有均质N末端的异源多肽的方法，其中所述异源多肽从表达的融合蛋白中自动蛋白水解切割，所述融合蛋白包含具有自身蛋白酶N的自身蛋白水解活性的多肽， 通过NOPO自身蛋白水解活性，瘟病毒和异源多肽的SUP> pro 。

公开(公告)号：US20060228783A1

公开(公告)日：2006-10-12

申请号：US10568337

申请日：2004-08-12

Applicant: Jorg Windisch ,  Kurt Schoergendorfer ,  Norbert Palma ,  Franz Knauseder ,  Hans Boehling

Inventor： Jorg Windisch ,  Kurt Schoergendorfer ,  Norbert Palma ,  Franz Knauseder ,  Hans Boehling

IPC: C12P21/04 ,  C07H21/04 ,  C12N15/74 ,  C12N1/21 ,  C07K14/56

CPC classification number: C12N15/70 ,  C07K14/56 ,  C07K2319/02 ,  C07K2319/034

Abstract: The present invention relates to an expression vector, comprising a polynucleotide encoding a fusion protein comprising the signal sequence of the gac gene of Pseudomonas diminuta and a polypeptide of interest, a prokaryotic host cell transformed with such an expression vector and a process for production of a polypeptide of interest using said host cell and said expression vector.

Abstract translation: 本发明涉及表达载体，其包含编码融合蛋白的多核苷酸，所述融合蛋白包含缺陷假单胞菌gac基因的信号序列和感兴趣的多肽，用这种表达载体转化的原核宿主细胞和生产 使用所述宿主细胞和所述表达载体的目标多肽。

公开(公告)号：US20050084969A1

公开(公告)日：2005-04-21

申请号：US10497015

申请日：2002-11-26

Applicant: Kurt Schorgendorfer ,  Jorg Windisch ,  Renate Kunert ,  Florian Unterluggauer

Inventor： Kurt Schorgendorfer ,  Jorg Windisch ,  Renate Kunert ,  Florian Unterluggauer

IPC: C12N15/09 ,  A61K38/22 ,  A61P7/00 ,  A61P7/06 ,  C07K14/505 ,  C12N5/10 ,  C12N15/85 ,  C12N15/90 ,  C12P21/02 ,  C12N5/06

CPC classification number: C07K14/505 ,  A61K2121/00 ,  C12N15/85 ,  C12N15/90 ,  C12N15/907 ,  C12N2840/203 ,  C12P21/02

Abstract: The invention relates to a method for producing a transformed eukaryotic host cell which expresses a recombinant polypeptide of interest which method comprises introducing into a eukaryotic host cell: (a) a first polynucleotide vector which comprises (i) a first nucleotide sequence which encodes a recombinant polypeptide of interest, and (ii) a second nucleotide sequence encoding a selectable marker, which second nucleotide sequence is amplified when the host cell is contacted with a selection agent; and (b) a second polynucleotide vector having essentially the same nucleotide sequence as the first polynucleotide vector except that the second nucleotide sequence is replaced with a third nucleotide sequence which encodes a different selectable marker; the first polynucleotide vector and second polynucleotide vector being stably integrated into the genome of the host cell.

Abstract translation: 本发明涉及用于产生表达重组感兴趣的多肽的转化真核宿主细胞的方法，所述方法包括将真核宿主细胞导入：（a）第一多核苷酸载体，其包含（i）第一核苷酸序列，其编码重组 多肽，和（ii）编码选择性标记的第二核苷酸序列，当宿主细胞与选择剂接触时，第二核苷酸序列被扩增; 和（b）第二多核苷酸载体，其具有与第一多核苷酸载体基本上相同的核苷酸序列，除了第二核苷酸序列被编码不同选择标记的第三核苷酸序列替代; 第一多核苷酸载体和第二多核苷酸载体稳定整合到宿主细胞的基因组中。