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    • 6. 发明授权
    • Recombinant DNA expression vectors and DNA compounds that encode
isopenicillin N synthetase from penicillium chrysogenum
    • 重组DNA表达载体和编码来自产黄青霉的异青霉素N合成酶的DNA化合物
    • US4892819A
    • 1990-01-09
    • US801523
    • 1985-11-25
    • Lucinda G. CarrThomas D. IngoliaStephen W. QueenerPaul L. Skatrud
    • Lucinda G. CarrThomas D. IngoliaStephen W. QueenerPaul L. Skatrud
    • C12N15/09C12N1/14C12N1/15C12N1/20C12N1/21C12N9/00C12N15/00C12N15/52C12N15/69C12N15/70C12N15/80C12N15/90C12P37/00C12R1/19C12R1/75C12R1/82
    • C12N9/93C12N15/69C12N15/70C12N15/80C12N15/90
    • The present invention comprises novel DNA compounds that encode isopenicillin N synthetase. The invention also comprises methods, transformants, and polypeptides related to the novel DNA compounds. The novel isopenicillin N synthetase-encoding DNA, together with its associated transcription and translation activating sequence, was isolated from Penicillium chrysogenum. The isopenicillin N synthetase-encoding DNA can be used to construct novel E. coli expression vectors that drive expression of isopenicillin N synthetase in E. coli. The intact P. chrysogenum isopenicillin N synthetase-encoding DNA and associated transcription and translation activating sequence can also be used to construct expression vectors that drive expression of the isopenicillin N synthetase in P. chrysogenum and Cephalosporium acremonium. The transcription and translation activating sequence can be fused to a hygromycin phosphotransferase-encoding DNA segment and placed onto expression vectors that function in P. chrysogenum and C. acremonium. The transcription termination and mRNA polyadenylation signals of the P. chrysogenum isopenicillin N synthetase can be used to increase ultimate expression of a product encoded on a recombinant DNA vector.
    • 本发明包括编码异青霉素N合成酶的新型DNA化合物。 本发明还包括与新型DNA化合物相关的方法,转化体和多肽。 编码新颖的异青霉素N合成酶的DNA,连同其相关的转录和翻译活化序列,从产黄青霉中分离出来。 编码异青霉素N合成酶的DNA可用于构建驱动大肠杆菌中异青霉素N合成酶表达的新型大肠杆菌表达载体。 完整的产黄青霉素异青霉素N合成酶编码DNA和相关的转录和翻译活化序列也可用于构建表达产物,其驱动产黄青霉和顶头孢霉的异青霉素N合成酶的表达。 转录和翻译激活序列可以与编码潮霉素磷酸转移酶的DNA片段融合,并置于表达载体中,该表达载体在产黄青霉和顶头孢霉中起作用。 产黄青霉素异青霉素N合成酶的转录终止和mRNA聚腺苷酸化信号可用于增加编码在重组DNA载体上的产物的最终表达。
    • 7. 发明授权
    • DNA for directing transcription and expression of structural genes
    • 用于引导转录和表达结构基因的DNA
    • US4559302A
    • 1985-12-17
    • US438070
    • 1982-11-01
    • Thomas D. Ingolia
    • Thomas D. Ingolia
    • C12N15/70C12N15/00C07H15/12C12N1/00C12N1/20C12P21/00C12P21/02
    • C12N15/70Y10S435/849Y10S435/886
    • The present invention relates to a novel transcriptional and translational activating sequence. The novel activating sequence can be either chemically synthesized or isolated on a 0.17 kb PstI-SacI restriction fragment from plasmid pKC203, a plasmid of E. coli JR225 (ATCC 31912). The activating sequence directs expression of the aminoglycoside acetyltransferase aac(3)IV and hygromycin phosphotransferase aph(4) genes present on plasmid pKC203. A series of expression vectors have been constructed in which the activating sequence directs the expression of beta-galactosidase or hygromycin phosphotransferase. These vectors can be readily modified and have been designed to facilitate the subsequent cloning and expression of any gene of research or commercial interest. The expression and cloning vectors have been transformed into E. coli and other host cells in which the activating sequence functions.
    • 本发明涉及一种新的转录和翻译活化序列。 新的活化序列可以在质粒pKC203(大肠杆菌JR225(ATCC 31912)的质粒)的0.17kb PstI-SacI限制性片段上化学合成或分离。 活化序列指示存在于质粒pKC203上的氨基糖苷酰乙酰转移酶aac(3)IV和潮霉素磷酸转移酶aph(4)基因的表达。 已经构建了一系列表达载体,其中活化序列指导β-半乳糖苷酶或潮霉素磷酸转移酶的表达。 这些载体可以容易地被修饰并被设计成有助于随后克隆和表达任何研究或商业利益的基因。 表达和克隆载体已经转化到大肠杆菌和其中启动序列起作用的其它宿主细胞。
    • 8. 发明授权
    • Recombinant DNA expression vectors and DNA compounds which encode
isopenicillin N synthetase
    • 编码异青霉素N合成酶的重组DNA表达载体和DNA化合物
    • US4885251A
    • 1989-12-05
    • US895008
    • 1986-08-08
    • Thomas D. IngoliaStephen W. QueenerSuellen M. SamsonPaul L. SkatrudOtis W. Godfrey
    • Thomas D. IngoliaStephen W. QueenerSuellen M. SamsonPaul L. SkatrudOtis W. Godfrey
    • C12N15/52C12N9/00C12N15/69C12N15/70C12N15/74C12N15/80
    • C12N15/80C12N15/69C12N9/93
    • The present invention comprises novel DNA compounds that encode isopenicillin N synthetase and also comprises related methods, transformants, and polypeptides. The novel isopenicillin N synthetase-encoding DNA, together with its associated transcriptional and translational activating sequence, was isolated from Cephalosporium acremonium and cloned into an E. coli cloning vector. The isopenicillin N synthetase-encoding DNA has been used to construct novel E. coli expression vectors that drive expression of a stable, active, and novel isopenicillin N synthetase in E. coli. The intact C. acremonium isopenicillin N synthetase-encoding DNA and associated transcriptional and translational activating sequence have also been used to construct C. acremonium expression vectors that drive expression of the isopenicillin N synthetase in C. acremonium. The C. acremonium transcriptional and translational activating sequence has further been fused to a hygromycin phosphotransferase-encoding DNA segment and placed onto C. acremonium expression vectors. Useful derivatives of the novel compounds and vectors are also described.
    • 本发明包括编码异青霉素N合成酶的新型DNA化合物,还包括相关方法,转化体和多肽。 编码新的异青霉素N合成酶DNA,连同其相关的转录和翻译活化序列,从头孢霉头孢菌分离并克隆到大肠杆菌克隆载体中。 编码异青霉素N合成酶的DNA已经用于构建新的大肠杆菌表达载体,其在大肠杆菌中驱动稳定的,活性的和新的异青霉素N合成酶的表达。 编码完整的顶头孢霉素异青霉素N合成酶的DNA和相关的转录和翻译活化序列也被用于构建顶头孢霉表达载体,其驱动顶头孢霉中异青霉素N合成酶的表达。 顶头孢霉转录和翻译激活序列进一步融合到编码潮霉素磷酸转移酶的DNA片段并置于顶头孢霉表达载体上。 还描述了新化合物和载体的有用衍生物。