Mitsui Toatsu Chemicals, Incorporated

Bridgestone Tire Company Limited

Mitsu Chemicals, Inc.

Yoshiyuki Nakajima

Nobuhiro Fukuhara

Setsuo Yoshino

Satori Sone

Nobuyoshi Makiguchi

Maki Suzuki

Kaoru Yamamoto

Mamoru Kadono

Tetsuji Saigusa

Tomohiro Koga

Tomoyuki Se

Amino acid sequence of L-phenylalanine ammonia-lyase

Escherichia coil 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 at a temperature 40.degree. C. or over so that the expression of said desired foreign gene was suppressed. This E. coil (i.e., transformant) was cultured at 40.degree. C. or over in a first process to suppress the expression of the foreign gene and to support sufficient cell growth and thereafter below 40.degree. C. 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.

Method of regulating expression of a foreign gene by controlling culture

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.

Method of regulating expression of a foreign gene by controlling the

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 at a temperature 40.degree. C. or over so that the expression of said desired foreign gene was suppressed. This E. coli (i.e., transformant) was cultured at 40.degree. C. or over in a first process to suppress the expression of the foreign gene and to support sufficient cell growth and thereafter below 40.degree. C. 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.

A recombinant plasmid for the expression of phenylalanine ammonia-lyase (PAL) is constructed by incorporating therein a combined promoter comprising (a) the fusion promoter (the tac promoter) composed of the trp promoter minus 35 region and the lac UV-5 promoter minus 10 region and (b) the P.sub.L promoter of the lambda phage, the tac promoter and the P.sub.L promoter being connected so as to have the same directional property. This recombinant plasmid permits more efficient expression of in Escherichia coli.

Recombinant plasmid for the expression of L-phenylalanine ammonia-lyase

A reinforced rubber hose mainly consisting of a hose body composed of inner and outer rubber layers and a reinforcing layer interposed therebetween and adapted to be secured or fastened at its two ends to a pair of cylindrical mouth pieces to be connected to the associated pipe ends. At least free end of the reinforcing layer is composed of inner and outer reinforcing layers. The inner reinforcing layer is wound around a clamping ring arranged at each end of the hose body from its inside toward the outside thereof to form a turn-back portion and the outer reinforcing layer is superimposed about and covering the turn-back portion of the inner reinforcing layer.

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Yokohama, Japan

Yoshiyuki Nakajima