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1. 发明授权

US4375514A Preparation and use of recombinant plasmids containing genes for alkaline phosphatases 失效
标题翻译：含有碱性磷酸酶基因的重组质粒的制备和使用
公开(公告)号：US4375514A
公开(公告)日：1983-03-01
申请号：US174316
申请日：1980-08-01
申请人： Gerhard Siewert , Werner Boidol , Joachim Daum
发明人： Gerhard Siewert , Werner Boidol , Joachim Daum
IPC分类号： C12N15/09 , C07H21/04 , C12N1/20 , C12N9/16 , C12N9/18 , C12P19/34 , C12R1/19 , C12R1/43 , C12N15/00 , C12N1/00
CPC分类号： C12N9/16 , C07K2319/00 , C07K2319/034 , Y10S435/82 , Y10S435/849 , Y10S435/881
摘要： PhoA mutant E. coli SB44, prepared by mutation of E. coli HB101 with an N-nitroso compound, can be used to identify and isolate recombinant plasmids into which a phoA gene has been incorporated. These plasmids can be used to transform bacteria which can be cloned and incubated to provide alkaline phosphatase in high yield. Moreover, these plasmid vectors can be modified in various ways so that the N-terminal amino acid sequence of phoA is followed in reading phase by the DNA coding for some other protein. In turn, these new plasmids can be used to transform bacteria which can be cloned and incubated to produce fusion proteins comprising the desired other protein in high yield and outside of the cell membrane in the periplasmatic space.
摘要翻译：通过用N-亚硝基化合物突变大肠杆菌HB101制备的PhoA突变型大肠杆菌SB44可用于鉴定并分离其中掺入phoA基因的重组质粒。这些质粒可用于转化可以克隆和培养的细菌，以高产率提供碱性磷酸酶。此外，这些质粒载体可以以各种方式进行修饰，使得phoA的N-末端氨基酸序列在编码一些其他蛋白质的DNA的阅读阶段被遵循。反过来，这些新的质粒可用于转化可以克隆和孵育的细菌，以产生高产量的包含期望的其它蛋白质的融合蛋白质，并且在周质空间内的细胞膜外。

2. 发明授权

US6114510A Process for the purification of recombinant human interleukin-8 失效
标题翻译：纯化重组人白介素-8的方法
公开(公告)号：US6114510A
公开(公告)日：2000-09-05
申请号：US77757
申请日：1998-06-11
申请人： Peter Scholz , Peter Donner , Joachim Daum , Werner Boidol , Andre Koltermann
发明人： Peter Scholz , Peter Donner , Joachim Daum , Werner Boidol , Andre Koltermann
IPC分类号： C12N15/09 , C07K1/36 , C07K14/54 , C12N1/21 , C12N15/24 , C12P21/02 , C12R1/19 , C07K1/14 , C07K1/16 , C07K1/18 , C07K14/00
CPC分类号： C07K1/36 , C07K14/5421
摘要： A process is described for the purification of recombinant interleukin-8, which allows the production of interleukin-8 under Good Manufacturing Practice (GMP) conditions.
摘要翻译： PCT No.PCT / EP96 / 05540 Sec。 371日期：1998年6月11日 102（e）1998年6月11日PCT 1996年12月11日PCT PCT。公开号WO97 / 21813 日期1996年6月19日描述了用于纯化重组白介素-8的方法，其允许在良好生产规范（GMP）条件下生产白细胞介素-8。

3. 发明授权

US4543329A Process for the specific cleavage of protein sequences from proteins 失效
标题翻译：蛋白质序列特异性切割的方法
公开(公告)号：US4543329A
公开(公告)日：1985-09-24
申请号：US395433
申请日：1982-07-06
申请人： Joachim Daum , Gerhard Siewert , Michael Topert , Hartmut Seliger
发明人： Joachim Daum , Gerhard Siewert , Michael Topert , Hartmut Seliger
IPC分类号： C07K14/575 , C07K14/61 , C07K14/62 , C12N15/62 , C12P21/06 , C12N15/00 , C12P21/02
CPC分类号： C07K14/62 , C07K14/57554 , C07K14/61 , C12N15/62 , C12P21/06 , C07K2319/00 , C07K2319/50 , C07K2319/61 , C07K2319/75 , Y10S930/31
摘要： A process for cleavage of a protein (a) sequence from a protein containing the tetrapeptide sequence Pro-Xyz-Gly-Pro, wherein Xyz can be any desired amino acid, and reacting the protein with a collagenase whereby the Xyz-Gly bond is selectively cleaved, thereby producing a protein with the sequence Gly-Pro at its N-terminal end; or (b) containing the sequence Met-Pro; and subsequently selectively cleaving the glycine or methionine residue from the N-terminal end of the product protein with an aminoacylproline aminopeptidase and selectively cleaving the proline residue therefrom with a proline aminopeptidase; or subsequently selectively cleaving the Gly-Pro or Met-Pro residue from the N-terminal end of the produced protein using a postproline dipeptidylaminopeptidase. In a special embodiment, the protein which is split contains the sequence -Zxy-Uvw-Pro- on the carboxy end of the sequence Pro-Xyz-Gly-Pro, wherein Zxy and Uvw are independently any natural amino acid in the genetic code except Pro, whereby after the collagenase reaction, there is produced a protein with the sequence Gly-Pro-Zxy-Uvw-Pro at the N-terminal end. This is split by subsequently selectively cleaving the Pro-Zxy-bond by treating the protein with postproline dipeptidylaminopeptidase (PPDA), thereby producing protein having the sequence Zxy-Uvw-Pro at its N-terminal end, followed by cleaving the Zxy acid from the resultant protein by treating the latter with leucine aminopeptidase.
摘要翻译：从含有四肽序列Pro-Xyz-Gly-Pro的蛋白质中切割蛋白质（a）序列的方法，其中Xyz可以是任何所需的氨基酸，并使蛋白质与胶原酶反应，由此Xyz-Gly键是选择性的从而在其N-末端产生具有Gly-Pro序列的蛋白质; 或（b）含有序列Met-Pro; 并随后用氨酰脯氨酸氨基肽酶选择性地从产物蛋白的N末端切割甘氨酸或甲硫氨酸残基，并用脯氨酸氨基肽酶选择性地切割脯氨酸残基; 或随后使用后脯氨酸二肽基氨基肽酶从产生的蛋白质的N末端选择性地切割Gly-Pro或Met-Pro残基。在一个特殊的实施方案中，分裂的蛋白质包含序列Pro-Xyz-Gly-Pro的羧基末端的序列-Zxy-Uvw-Pro，其中Zxy和Uvw独立地是遗传密码中的任何天然氨基酸， Pro，由此在胶原酶反应之后，在N末端产生具有序列Gly-Pro-Zxy-Uvw-Pro的蛋白质。通过用后脯氨酸二肽基氨基肽酶（PPDA）处理蛋白质随后选择性地切割Pro-Zxy-键，从而在其N-末端产生具有序列Zxy-Uvw-Pro的蛋白质，随后将Zxy酸从通过用亮氨酸氨基肽酶处理后者产生的蛋白质。

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