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    Method of regulating expression of a foreign gene by controlling the sugar concentration in a medium and a process of producing a foreign product thereby
    4.
    发明授权
    Method of regulating expression of a foreign gene by controlling the sugar concentration in a medium and a process of producing a foreign product thereby 失效
    通过控制培养基中的糖浓度和由此制造异物的方法来调节外源基因的表达的方法

    公开(公告)号：US5139935A

    公开(公告)日：1992-08-18

    申请号：US659472

    申请日：1991-02-25

    申请人： Nobuhiro Fukuhara ,  Setsuo Yoshino ,  Satori Sone ,  Yoshiyuki Nakajima ,  Nobuyoshi Makiguchi

    发明人： Nobuhiro Fukuhara ,  Setsuo Yoshino ,  Satori Sone ,  Yoshiyuki Nakajima ,  Nobuyoshi Makiguchi

    IPC分类号： C12N1/21 ,  C12N9/88 ,  C12N15/60 ,  C12N15/70

    CPC分类号： C12N15/70 ,  C12N9/88

    摘要： Escherichia coli carrying a hybrid plasmid having been constructed by inserting a desired foreign gene into an expression vector so as to permitting expression of said desired foreign gene therein was cultured in a medium containing a sugar component utilizable by the E. coli as a carbon source at a concentration of 0.3% or more so that the expression of said desired foreign gene was suppressed. This E. coli (i.e., transformant) was cultured in the medium in which the sugar concentration was maintained at 0.3% or more in a first process so as to supress the suppression of the foreign gene and to support sufficient cell growth and thereafter at less than 0.3% in a second process to release the suppression of the expression so as to permit effective production of the foreign gene product, which resulted in high concentration of the foreign gene product in the final culture.

    摘要翻译： 携带通过将期望的外源基因插入到表达载体中以便允许其中表达所需外源基因的杂交质粒的大肠杆菌在含有作为碳源的大肠杆菌可利用的糖成分的培养基中培养 浓度为0.3％以上，能够抑制所希望的外源基因的表达。 将该大肠杆菌（即转化体）在第一种方法中将糖浓度保持在0.3％以上的培养基中培养，以抑制外源基因的抑制并支持足够的细胞生长，此后在少于 在第二过程中超过0.3％以释放表达的抑制，以便有效地产生外源基因产物，这导致外源基因产物在最终培养物中的高浓度。

    Amino acid sequence of L-phenylalanine ammonia-lyase
    6.
    发明授权
    Amino acid sequence of L-phenylalanine ammonia-lyase 失效
    L-苯丙氨酸氨裂解酶的氨基酸序列

    公开(公告)号：US06472196B1

    公开(公告)日：2002-10-29

    申请号：US08214018

    申请日：1994-03-15

    申请人： Nobuhiro Fukuhara ,  Setsuo Yoshino ,  Kaoru Yamamoto ,  Tomoyuki Se ,  Satori Sone ,  Yoshiyuki Nakajima ,  Maki Suzuki ,  Nobuyoshi Makiguchi

    发明人： Nobuhiro Fukuhara ,  Setsuo Yoshino ,  Kaoru Yamamoto ,  Tomoyuki Se ,  Satori Sone ,  Yoshiyuki Nakajima ,  Maki Suzuki ,  Nobuyoshi Makiguchi

    IPC分类号： C12N988

    CPC分类号： C12P13/222 ,  C12N9/88

    摘要： In order to provide technical information necessary for the production of L-phenylalanine ammonia-lyase (PAL) by utilizing genetic engineering techniques, the structural gene for PAL and the amino acid sequence of PAL have been elucidated in Rhodosporidium toruloides, and novel recombinant DNA plasmids (e.g., pSW101, pYtrp6 and pKY201) have been created by inserting a DNA strand coding for the PAL gene between the 3′-terminus of the promoter region and the 5′-terminus of the terminator region. Moreover, transformants having such a novel recombinant DNA plasmid have been created, and a process for the production of PAL by growing such a novel transformant so as to cause PAL to be produced and accumulated in the culture has been established. Furthermore, there has been established a novel technique for the production of L-phenylalanine by reacting an ammonia donor with cinnamic acid in the presence of the PAL prepared by the aforesaid novel process.

    摘要翻译： 为了提供利用遗传工程技术生产L-苯丙氨酸氨裂解酶（PAL）所必需的技术信息，已经在Rhodosporidium toruloides中阐明了PAL的结构基因和PAL的氨基酸序列，新的重组DNA质粒 （例如，pSW101，pYtrp6和pKY201）是通过将编码PAL基因的DNA链插入到启动子区域的3'端和终止区域的5'末端而产生的。此外，具有这样一种新颖的转化体 已经建立了重组DNA质粒，并且通过生长这样一种新的转化体以使培养中产生和积累PAL来生产PAL的方法已被建立。此外，已经建立了一种生产新技术 在通过上述新方法制备的PAL存在下，使氨供体与肉桂酸反应，生成L-苯丙氨酸。