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1.发明授权Transcription terminator and related recombinant DNA vectors and transformants 失效转录终止子和相关重组DNA载体和转化体
公开(公告)号：US5093252A
公开(公告)日：1992-03-03
申请号：US515538
申请日：1990-04-30
摘要： A transcription terminator is disclosed comprising a DNA sequence capable of functioning as a transcription terminator in a microorganism of the order Actinomycetales. These transcription terminators are functional in both gram-positive and gram-negative microorganisms, especially Streptomyces ambofaciens. Additionally, a series of recombinant DNA cloning vectors suitable for isolating the disclosed transcription terminators are set forth.
摘要翻译： 公开了一种转录终止子，其包含能够作为放线菌纲的微生物中的转录终止子的DNA序列。 这些转录终止子在革兰氏阳性和革兰氏阴性微生物，特别是链霉菌（Streptomyces ambofaciens）中均有功能。 另外，阐述了一系列用于分离所公开的转录终止子的重组DNA克隆载体。
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2.发明授权Method of using bacteriophage lambda p.sub.1 promoter to produce a functional polypeptide in streptomyces 失效使用噬菌体λp1启动子在链霉菌中产生功能性多肽的方法
公开(公告)号：US4766066A
公开(公告)日：1988-08-23
申请号：US655181
申请日：1984-09-27
IPC分类号： C12N15/09 , C12N1/20 , C12N9/00 , C12N15/73 , C12N15/76 , C12P21/00 , C12R1/465 , C12N15/00 , C12P19/34
CPC分类号： C12N15/73 , C12N15/76 , Y10S435/886 , Y10S435/889 , Y10S435/896
摘要： The invention relates to a method and cloning vehicle for the expression of a functional polypeptide in Streptomyces. A recombinant DNA cloning vehicle was genetically engineered to bring the expression of the neomycin phosphotransferase gene under the control of the Escherichia coli bacteriophage .lambda.p.sub.L promoter.
摘要翻译： 本发明涉及用于在链霉菌中表达功能性多肽的方法和克隆载体。 基因工程改造重组DNA克隆载体，使新霉素磷酸转移酶基因的表达在大肠杆菌噬菌体λpL启动子的控制下。
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公开(公告)号：US5190871A
公开(公告)日：1993-03-02
申请号：US364959
申请日：1989-06-12
申请人： Karen L. Cox , Stuart A. Kuhstoss , R. Nagaraja Rao , Mark A. Richardson , Brigitte E. Schoner , Eugene T. Seno
发明人： Karen L. Cox , Stuart A. Kuhstoss , R. Nagaraja Rao , Mark A. Richardson , Brigitte E. Schoner , Eugene T. Seno
摘要： The present invention provides a method for transforming an actinomycete with an integrating vector which has the advantages of high transformation rates into a broad host range, site-specific integration, and stable maintenance without antibiotic selection. Also provided are methods for the increased production of antibiotics and for the production of hybrid antibiotics.
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公开(公告)号：US4710464A
公开(公告)日：1987-12-01
申请号：US655184
申请日：1984-09-27
IPC分类号： C12N15/67 , C12N15/70 , C12N15/76 , C12P19/34 , C07H21/04 , C12N1/00 , C12N1/16 , C12N1/18 , C12N1/20 , C12N15/00 , C12P21/00
摘要： The present transcription terminators are DNA sequences characterized by their ability to terminate the transcription of DNA sequences. The present sequences are portable, have a variety of restriction endonuclease sites and are useful in controlling the expression in hosts of DNA sequences coding for prokaryotic, eukaryotic and viral proteins and polypeptides.
摘要翻译： 本转录终止子是以其终止DNA序列转录的能力为特征的DNA序列。 本发明的序列是便携式的，具有多种限制性内切核酸酶位点，并且可用于控制编码原核，真核和病毒蛋白质和多肽的DNA序列的宿主中的表达。
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公开(公告)号：US4935340A
公开(公告)日：1990-06-19
申请号：US742349
申请日：1985-06-07
IPC分类号： C12N15/09 , C12N1/20 , C12N1/21 , C12N15/52 , C12N15/76 , C12Q1/68 , C12R1/03 , C12R1/045 , C12R1/07 , C12R1/15 , C12R1/29 , C12R1/365 , C12R1/465 , C12R1/625 , C12R1/745 , C12R1/80 , G01N33/50
摘要： The present invention is a method for isolating antibiotic biosynthetic genes. To practice the method, an antibiotic resistance-conferring DNA segment is labelled and used as a probe to find, via DNA hydridization, homologous DNA in a genetic library which comprises chromosomal and plasmid DNA of an antibiotic-producing organism. Individual vectors of the genetic library which hybridize to the antibiotic resistance-conferring gene, and which comprise .about.1-45 kb of contiguous DNA from the antibiotic-producing organism, which also comprise an antibiotic biosynthetic gene. The present method is exemplified by using the erythromycin resistance-conferring gene of Streptomyces erythreus to clone the erythromycin biosynthetic pathway from the same organism. The erythromycin biosynthetic pathway isolated with the present method synthesizes erythromycin when introduced into S. lividans TK23.
摘要翻译： 本发明是分离抗生素生物合成基因的方法。 为了实施该方法，将抗生素抗性赋予DNA片段标记并用作探针，通过DNA杂交在包含产生抗生素的生物体的染色体和质粒DNA的遗传文库中发现同源DNA。 与抗生素耐药基因杂交的基因文库的单个载体，其包含来自产生抗生素的生物体的差异1-45kb的连续DNA，其也包含抗生素生物合成基因。 本发明的方法通过使用红霉素抗性赋予基因，从相同的生物体克隆红霉素生物合成途径。 用本发明方法分离的红霉素生物合成途径将红霉素引入青光眼链霉菌TK23后合成红霉素。
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公开(公告)号：US4960704A
公开(公告)日：1990-10-02
申请号：US205011
申请日：1988-05-31
CPC分类号： C07K14/62 , C07K14/57581 , C12N15/62 , C12N15/81 , C12N15/8209 , C12N9/12 , C07K2319/00 , C07K2319/02
摘要： A modified hygromycin B resistance-conferring gene either alone or in translational reading phase with a gene or portion of a gene is disclosed. The invention further comprises recombinant DNA cloning vectors and transformants of the aforementioned DNA.
摘要翻译： 公开了改良的潮霉素B抗性基因，其单独或翻译阅读阶段与基因或基因的一部分。 本发明还包括重组DNA克隆载体和上述DNA的转化体。
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7.发明授权Novel plasmid shuttle vectors conferring spiramycin resistance in streptomyces 失效新型质粒穿梭载体赋予链霉菌中的螺旋霉素抗性
公开(公告)号：US4904598A
公开(公告)日：1990-02-27
申请号：US841981
申请日：1986-03-20
申请人： R. Nagaraja Rao
发明人： R. Nagaraja Rao
CPC分类号： C12N15/65 , C12N15/76 , Y10S435/886
摘要： A novel gene conferring resistance to spiramycin in Streptomyces griseofuscus was cloned from a genomic library of Streptomyces ambofaciens DNA. An eighteen kb fragment of S. ambofaciens DNA surrounding the spiramycin resistance gene was isolated from this library on a plasmid cosmid designated pKC514. The novel spiramycin-resistance gene can be isolated on an .about.3.4 kb Sau3AI fragment by subcloning restriction fragments obtained from the pKC514 insert DNA. This Sau3AI fragment contains all of the information required for the expression of the spiramycin resistant phenotype in Streptomyces. Vectors and transformants containing the novel spiramycin resistance gene are provided.
摘要翻译： 从链霉菌（Streptomyces ambofaciens）DNA的基因组文库中克隆了赋予灰霉菌链霉菌中螺旋霉素抗性的新基因。 在名为pKC514的质粒粘粒上，从该文库中分离出围绕螺旋霉素抗性基因的淀粉芽孢杆菌DNA的十八分之一片段。 通过从pKC514插入DNA获得的亚克隆限制性片段，可以在DIFFERENCE 3.4kb Sau3AI片段上分离新的螺旋霉素抗性基因。 该Sau3AI片段含有在链霉菌中表达螺旋霉素抗性表型所需的所有信息。 提供了含有新型螺旋霉素抗性基因的载体和转化体。
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8.发明授权
公开(公告)号：US5098837A
公开(公告)日：1992-03-24
申请号：US203387
申请日：1988-06-07
CPC分类号： C12P19/62 , C12N15/52 , C12N15/76 , Y10S435/886
摘要： Spiramycin antibiotic biosynthetic genes of Streptomyces ambofaciens are provided by the present invention, in addition to a variety of recombinant DNA vectors. The genes also function in other macrolide producing organisms. The genes can be used to increase or otherwise alter the macrolide antibiotic-producing ability of an organism. The present invention also provides host strains comprising mutant spiramycin biosynthetic genes which can be used to generate novel antibiotics. Also provided is a method for preparing the mutant gene comprising mutating cloned spiramycin biosynthetic DNA by transposon mutagenesis with subsequent transformation into a macrolide-antibiotic producing host and homologous recombination into its genome, to generate stable mutant cell lines.
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公开(公告)号：US4921801A
公开(公告)日：1990-05-01
申请号：US842102
申请日：1986-03-20
CPC分类号： C12N15/76 , Y10S435/849 , Y10S435/889
摘要： Novel recombinant DNA cosmid shuttle vectors and a method of using them in the construction of genomic DNA libraries are described. The vectors demonstrate the incorporation of both the size selection and in vitro packaging mechanisms of lambda into a Streptomyces-E. coli shuttle vector by the incorporation of two or more COS sequences of bacteriophage lambda.
摘要翻译： 描述了新的重组DNA粘粒穿梭载体及其在构建基因组DNA文库中的使用方法。 这些载体证明将λ的大小选择和体外包装机制纳入Streptomyces-E中。 大肠杆菌穿梭载体通过结合两个或多个COS序列的噬菌体λ。
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公开(公告)号：US4843002A
公开(公告)日：1989-06-27
申请号：US655180
申请日：1984-09-27
CPC分类号： C12N15/76 , C12N15/65 , C12N9/1029 , Y10S435/872 , Y10S435/886 , Y10S435/889 , Y10S435/896
摘要： A novel method of selecting Streptomyces recombinant DNA-containing host cells and vectors useful in exemplifying the method are described. The vectors confer apramycin resistance to sensitive Streptomyces host cells and thus provide a convenient method of selecting Streptomyces transformants. The apramycin resistance-conferring gene used in the method is an acetyltransferase aac(3)IV gene and can be isolated from E. coli K12 BE1041/pKC309 (NRRL B-15827) on an .about.1.5 kb PstI-EcoRI restriction fragment.
摘要翻译： 描述了选择含有重链DNA的宿主细胞的新方法和用于举例说明该方法的载体。 这些载体赋予了对敏感链霉菌宿主细胞的安普霉素抗性，从而提供了选择链霉菌转化体的方便的方法。 该方法中使用的安普霉素抗性基因是乙酰转移酶aac（3）IV基因，并且可以在DIFFERENCE 1.5kb PstI-EcoRI限制性片段上从大肠杆菌K12 BE1041 / pKC309（NRRL B-15827）中分离。
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