JP2002539793A5 - - Google Patents

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JP2002539793A5
JP2002539793A5 JP2000606759A JP2000606759A JP2002539793A5 JP 2002539793 A5 JP2002539793 A5 JP 2002539793A5 JP 2000606759 A JP2000606759 A JP 2000606759A JP 2000606759 A JP2000606759 A JP 2000606759A JP 2002539793 A5 JP2002539793 A5 JP 2002539793A5
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【特許請求の範囲】
【請求項1】 ポリペプチドを製造する方法であって、
(a)菌類宿主細胞を、ポリペプチドの製造を助ける培地中で培養し、菌類宿主細胞は、第1の核酸配列に対して外来のプロモーターを含む第2の核酸配列に機能しうる形で連結された、ポリペプチドをコードする第1の核酸配列を含み、かつ、前記プロモーターは、配列番号:1のヌクレオチド1〜3949、配列番号:2のヌクレオチド1〜938および配列番号:3のヌクレオチド1〜3060並びにそのサブ配列;並びに、その変異体、ハイブリッドおよび縦列プロモーターからなる群より選択される配列を含み;そして
(b)培養培地からポリペプチドを分離する;
ことを含んで成る方法。
【請求項2】 前記プロモーターが、大腸菌(E. coli)NRRL B-30067に含有されるプラスミドpECO3;大腸菌(E. coli)NRRL B-30071に含有されるプラスミドpFAMG;または大腸菌(E. coli)NRRL B-30075に含有されるプラスミドpQUINNに含まれる核酸配列を含む請求項1記載の方法。
【請求項3】 配列番号:1のヌクレオチド1〜3949、配列番号:2のヌクレオチド1〜938および配列番号:3のヌクレオチド1〜3060並びにそのサブ配列;ならびに、その変異体、ハイブリッドおよび縦列プロモーターからなる群より選択される核酸配列を含むプロモーター。
【請求項4】 大腸菌(E. coli)NRRL B-30067に含有されるプラスミドpECO3;大腸菌(E. coli)NRRL B-30071に含有されるプラスミドpFAMG;または大腸菌(E. coli)NRRL B-30075に含有されるプラスミドpQUINNに含まれる核酸配列を含む請求項3記載の単離されたプロモーター。
【請求項5】 請求項3記載のプロモーターに機能しうる形で連結された、ポリペプチドをコードする核酸配列を含む核酸構築物。
【請求項6】 請求項5記載の核酸構築物を含む組換え発現ベクター。
【請求項7】 請求項5記載の核酸構築物を含む組換え宿主細胞。
【請求項8】 変異体プロモーターを得るための方法であって、
(a)低いストリンジェンシー条件下でDNAを、(i)配列番号:1、配列番号:2もしくは配列番号:3、(ii)配列番号:1、配列番号:2もしくは配列番号:3に含まれるcDNA配列、(iii)前記(i)もしくは(ii)のサブ配列、または(iv)前記(i)、(ii)もしくは(iii)の相補鎖を含む核酸配列を用いてハイブリダイズし;そして
(b)DNAから変異体プロモーターを分離する;
ことを含んで成る方法。
【請求項9】 変異体プロモーターを得るための方法であって、(a)配列番号:1のヌクレオチド1〜3949、配列番号:2のヌクレオチド1〜938または配列番号:3のヌクレオチド1〜3060のプロモーターに、少なくとも1つの変異を導入し、ここで変異体プロモーターは、配列番号:1のヌクレオチド1〜3949、配列番号:2のヌクレオチド1〜938または配列番号:3のヌクレオチド1〜3060のプロモーターと同じプロモーター活性を有し;そして(b)変異体プロモーターを単離することを含む方法。
【請求項10】 (a)配列番号:4のアミノ酸22〜581、配列番号:5のアミノ酸19〜200または配列番号:6のアミノ酸1〜187と少なくとも65%の同一性を有するアミノ酸配列を含むポリペプチドをコードする核酸配列;
(b)配列番号:1のヌクレオチド4013〜5743、配列番号:2のヌクレオチド993〜1593または配列番号:3のヌクレオチド3061〜3678と少なくとも65%の相同性を有する核酸配列;
(c)中間の、中〜高い、高い、または非常に高いストリンジェンシー条件下で、(i)配列番号:1のヌクレオチド4013〜5743、配列番号:2のヌクレオチド993〜1593もしくは配列番号:3のヌクレオチド3061〜3678;(ii)配列番号:1のヌクレオチド4013〜5743、配列番号:2のヌクレオチド993〜1593もしくは配列番号:3のヌクレオチド3061〜3678に含まれるcDNA配列、または(iii)前記(i)もしくは(ii)の相補鎖とハイブリダイズする核酸配列;
(d)前記(a)、(b)または(c)の対立遺伝子変異体;ならびに
(e)活性を保持するポリペプチドをコードする、(a)、(b)、(c)または(d)のサブ配列;
からなる群より選択される核酸配列を有する核酸。
【請求項11】 大腸菌(E. coli)NRRL B-30067に含有されるプラスミドpECO3;大腸菌(E. coli)NRRL B-30071に含有されるプラスミドpFAMG;または大腸菌(E. coli)NRRL B-30075に含有されるプラスミドpQUINNに含まれる核酸配列を含む請求項10記載の核酸。
【請求項12】 適当な発現宿主中でポリペプチドの製造を指令する、1つ以上の制御配列に機能しうるように連結された、請求項10記載の核酸を含む核酸構築物。
【請求項13】 請求項12記載の核酸構築物を含む組換え発現ベクター。
【請求項14】 請求項12記載の核酸構築物を含む組換え宿主細胞。
【請求項15】 請求項10記載の核酸によってコードされるポリペプチド。
JP2000606759A 1999-03-22 2000-03-22 菌類細胞中で遺伝子を発現させるためのプロモーター Expired - Fee Related JP4620253B2 (ja)

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US27444999A 1999-03-22 1999-03-22
US09/274,449 1999-03-22
PCT/US2000/007815 WO2000056900A2 (en) 1999-03-22 2000-03-22 Promoter sequences derived from fusarium venenatum and uses thereof

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EP (2) EP2278016B1 (ja)
JP (4) JP4620253B2 (ja)
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