公开(公告)号：US20100120154A1

公开(公告)日：2010-05-13

申请号：US12532205

申请日：2008-03-20

申请人： Hesselien Touw-Riel ,  Marco Alexander Berg, Van Den

发明人： Hesselien Touw-Riel ,  Marco Alexander Berg, Van Den

IPC分类号： C12N15/85 ,  C12N15/82 ,  C12N15/80

CPC分类号： C12N15/80 ,  C07K14/385 ,  C12N15/1058 ,  C12N15/902 ,  C12N2800/30

摘要： The present invention discloses a method to construct eukaryotic cells having a target sequence in a chromosomal DNA sequence replaced by a replacement sequence of interest comprising: modifying a parent eukaryotic cell with a preference for NHR to provide a eukaryotic cell having an increased HR/NHR ratio as compared to the parent cell, providing two sets of DNA molecules of which the first set comprises DNA molecules each comprising a first non-functional fragment of the replacement sequence of interest flanked at its 5′-side by a DNA sequence substantially homologous to a sequence of the chromosomal DNA flanking the target sequence and the second set comprises DNA molecules each comprising a second non-functional fragment of the DNA replacement sequence of interest overlapping with the first non-functional fragment and flanked at its 3′-side by a DNA sequence substantially homologous to a sequence of the chromosomal DNA flanking the target sequence, wherein the first and second non-functional fragments become functional upon recombination; transforming the modified eukaryotic cells with both sets of DNA molecules; and growing the cells to obtain transformed progeny cells having a DNA molecule from the first set recombined with a DNA molecule from the second set into a DNA molecule comprising a functional replacement sequence and having the target sequence replaced by the functional replacement sequence.

摘要翻译： 本发明公开了一种构建真核细胞的方法，所述真核细胞在染色体DNA序列中被替换的目标序列替代，所述染色体DNA序列被替换的目标序列所替代，包括：以优选NHR修饰亲本真核细胞以提供具有增加的HR / NHR比的真核细胞 与亲本细胞相比，提供两组DNA分子，其中第一组包含DNA分子，每个DNA分子包含在其5'侧两侧的替换序列的第一个非功能性片段，其基本上与 在靶序列侧翼的染色体DNA的序列和第二组包含DNA分子，每个DNA分子包含与第一非功能片段重叠的DNA置换序列的第二个非功能性片段，并在其3'侧通过DNA侧接 基本上与靶序列侧翼的染色体DNA的序列同源的序列，其中第一和第二 非功能性片段在重组时变得有功能; 用两组DNA分子转化修饰的真核细胞; 并生长细胞以获得具有来自第一组的DNA分子的转化后代细胞与来自第二组的DNA分子重组为包含功能性替换序列的DNA分子并且将靶序列替换为功能性替换序列。

公开(公告)号：US20140154807A1

公开(公告)日：2014-06-05

申请号：US14146858

申请日：2014-01-03

申请人： Hesselien TOUW-RIEL ,  Marco Alexander BERG VAN DEN

发明人： Hesselien TOUW-RIEL ,  Marco Alexander BERG VAN DEN

IPC分类号： C12N15/80

CPC分类号： C12N15/80 ,  C07K14/385 ,  C12N15/1058 ,  C12N15/902 ,  C12N2800/30

摘要： The present invention discloses a method to construct eukaryotic cells having a target sequence in a chromosomal DNA sequence replaced by a replacement sequence of interest comprising: modifying a parent eukaryotic cell with a preference for NHR to provide a eukaryotic cell having an increased HR/NHR ratio as compared to the parent cell, providing two sets of DNA molecules of which the first set comprises DNA molecules each comprising a first non-functional fragment of the replacement sequence of interest flanked at its 5′-side by a DNA sequence substantially homologous to a sequence of the chromosomal DNA flanking the target sequence and the second set comprises DNA molecules each comprising a second non-functional fragment of the DNA replacement sequence of interest overlapping with the first non-functional fragment and flanked at its 3′-side by a DNA sequence substantially homologous to a sequence of the chromosomal DNA flanking the target sequence, wherein the first and second non-functional fragments become functional upon recombination; transforming the modified eukaryotic cells with both sets of DNA molecules; and growing the cells to obtain transformed progeny cells having a DNA molecule from the first set recombined with a DNA molecule from the second set into a DNA molecule comprising a functional replacement sequence and having the target sequence replaced by the functional replacement sequence.

摘要翻译： 本发明公开了一种构建真核细胞的方法，所述真核细胞在染色体DNA序列中被替换的目标序列替代，所述染色体DNA序列被替换的目标序列所替代，包括：以优选NHR修饰亲本真核细胞以提供具有增加的HR / NHR比的真核细胞 与亲本细胞相比，提供两组DNA分子，其中第一组包含DNA分子，每个DNA分子包含在其5'侧两侧的替换序列的第一个非功能性片段，其基本上与 在靶序列侧翼的染色体DNA的序列和第二组包含DNA分子，每个DNA分子包含与第一非功能片段重叠的DNA置换序列的第二个非功能性片段，并在其3'侧通过DNA侧接 基本上与靶序列侧翼的染色体DNA的序列同源的序列，其中第一和第二 非功能性片段在重组时变得有功能; 用两组DNA分子转化修饰的真核细胞; 并生长细胞以获得具有来自第一组的DNA分子的转化后代细胞与来自第二组的DNA分子重组为包含功能性替换序列的DNA分子并且将靶序列替换为功能性替换序列。

公开(公告)号：US20120276567A1

公开(公告)日：2012-11-01

申请号：US13505697

申请日：2010-11-04

申请人： Alrik Pieter Los ,  Brenda Vonk ,  Marco Alexander Berg Van Den ,  Robbertus Antonius Damveld ,  Cornelis Maria Jacobus Sagt ,  Adrianus Wilhelmus Hermanus Vollebregt ,  Margot Elisabeth Francoise Schooneveld-Bergmans

发明人： Alrik Pieter Los ,  Brenda Vonk ,  Marco Alexander Berg Van Den ,  Robbertus Antonius Damveld ,  Cornelis Maria Jacobus Sagt ,  Adrianus Wilhelmus Hermanus Vollebregt ,  Margot Elisabeth Francoise Schooneveld-Bergmans

IPC分类号： C12N1/15 ,  C12Q1/48 ,  C12N9/42 ,  C12N9/24 ,  C12N9/02 ,  C12N9/88 ,  C12N9/20 ,  C12N9/18 ,  C12N9/26 ,  C12N9/34 ,  C12N9/38 ,  C12N9/40 ,  C12N9/08 ,  C12N9/22 ,  C12P19/14 ,  C12P7/14 ,  C12P7/16 ,  C12P7/04 ,  C12P13/04 ,  C12P7/56 ,  C12P5/02 ,  C12N15/80 ,  C12P1/00 ,  C12P1/02 ,  C12Q1/34

CPC分类号： C12N15/80 ,  C12N9/2437 ,  C12N9/2445 ,  C12N9/2477 ,  C12P21/02 ,  C12Y302/01004 ,  C12Y302/01021 ,  C12Y302/01091 ,  Y02E50/343

摘要： The invention relates to a Talaromyces transformant comprising one or more recombinant gene, capable of producing cellulase in the absence of cellulase inducer in a glucose medium, having a cellulase activity of 2 WSU/ml or more, in 16 times or more diluted supernatant or broth.

摘要翻译： 本发明涉及一种含有一种或多种重组基因的塔拉霉菌转化体，其在16倍或更多稀释的上清液或肉汤中能够在葡萄糖培养基中不存在纤维素酶诱导物的情况下产生纤维素酶活性为2WUU / ml以上的纤维素酶 。