公开(公告)号：US20050079618A1

公开(公告)日：2005-04-14

申请号：US10841125

申请日：2004-05-07

申请人： Donald Court ,  Xin-tian Li ,  Jian-Dong Huang ,  Nina Costantino ,  Depei Liu

发明人： Donald Court ,  Xin-tian Li ,  Jian-Dong Huang ,  Nina Costantino ,  Depei Liu

IPC分类号： C12N1/20 ,  C12N15/70 ,  C12N15/74

CPC分类号： C12N1/20 ,  C12N15/70

摘要： Methods are disclosed herein for inducing homologous recombination in a host cell comprising a target nucleic acid, using a single-stranded nucleic acid molecule. The single-stranded nucleic acid molecule has a sufficient number of nucleotides homologous to the target nucleic acid to enable homologous recombination with the target nucleic acid. The host cell includes a de-repressible promoter operably linked to a nucleic acid encoding a single-stranded binding protein and is deficient for mismatch repair. Isolated host cells of use in this method are also disclosed.

摘要翻译： 本文公开了使用单链核酸分子在包含靶核酸的宿主细胞中诱导同源重组的方法。 单链核酸分子具有与目标核酸同源的足够数量的核苷酸，以实现与靶核酸的同源重组。 宿主细胞包括可操作地连接到编码单链结合蛋白的核酸并且缺陷以用于错配修复的去阻抑性启动子。 还公开了在该方法中使用的分离的宿主细胞。

公开(公告)号：US07521242B2

公开(公告)日：2009-04-21

申请号：US10841125

申请日：2004-05-07

申请人： Donald L. Court ,  Xin-tian Li ,  Jian-Dong Huang ,  Nina Costantino ,  Depei Liu

发明人： Donald L. Court ,  Xin-tian Li ,  Jian-Dong Huang ,  Nina Costantino ,  Depei Liu

IPC分类号： C12N15/87 ,  C12N15/90 ,  C12N1/21

CPC分类号： C12N1/20 ,  C12N15/70

摘要： Methods are disclosed herein for inducing homologous recombination in a host cell comprising a target nucleic acid, using a single-stranded nucleic acid molecule. The single-stranded nucleic acid molecule has a sufficient number of nucleotides homologous to the target nucleic acid to enable homologous recombination with the target nucleic acid. The host cell includes a de-repressible promoter operably linked to a nucleic acid encoding a single-stranded binding protein and is deficient for mismatch repair. Isolated host cells of use in this method are also disclosed.

摘要翻译： 本文公开了使用单链核酸分子在包含靶核酸的宿主细胞中诱导同源重组的方法。 单链核酸分子具有与目标核酸同源的足够数量的核苷酸，以实现与靶核酸的同源重组。 宿主细胞包括可操作地连接到编码单链结合蛋白的核酸并且缺陷以用于错配修复的去阻抑性启动子。 还公开了在该方法中使用的分离的宿主细胞。

公开(公告)号：US09222093B2

公开(公告)日：2015-12-29

申请号：US13534774

申请日：2012-06-27

申请人： Jian-Dong Huang ,  Ye Jin

发明人： Jian-Dong Huang ,  Ye Jin

IPC分类号： C12N15/113 ,  C12N15/63 ,  C12N15/67 ,  C12N15/72 ,  C12N15/115

CPC分类号： C12N15/63 ,  C12N15/115 ,  C12N15/635 ,  C12N15/67 ,  C12N15/72 ,  C12N2310/16 ,  C12N2310/3519 ,  C12N2320/10

摘要： A regulatable gene expression construct comprising a nucleic acid molecule comprising a two-way riboswitch operably linked to a target sequence. Also provided is a library screening strategy for efficient creation of target-specific riboswitches. A theophylline-repressible and IPTG-inducible riboswitch device achieves portable control of gene expression control in a ‘two-way’ manner. The default state of target genes is ON; the targets are switched off by adding theophylline, and switched back to the ON-state by adding IPTG without changing growth medium. The riboswitch device regulates gene expression in a portable, adjustable, and two-way manner with a variety of scientific and biotechnological applications.

摘要翻译： 一种可调节基因表达构建体，其包含核酸分子，其包含可操作地连接到靶序列的双向核糖开关。 还提供了一种用于有效创建目标特异性核糖开关的图书馆筛选策略。 茶碱抑制性和IPTG诱导型核糖开关器件以“双向”方式实现基因表达控制的便携式控制。 目标基因的默认状态为ON; 通过添加茶碱来关闭目标，并通过添加IPTG而不改变生长培养基而切换回ON状态。 核糖开关器件通过各种科学和生物技术应用以便携式，可调节和双向的方式调节基因表达。

公开(公告)号：US20130004980A1

公开(公告)日：2013-01-03

申请号：US13534774

申请日：2012-06-27

申请人： Jian-Dong Huang ,  Ye Jin

发明人： Jian-Dong Huang ,  Ye Jin

IPC分类号： C12N15/113 ,  C40B40/06 ,  C12Q1/34 ,  C07K17/10

CPC分类号： C12N15/63 ,  C12N15/115 ,  C12N15/635 ,  C12N15/67 ,  C12N15/72 ,  C12N2310/16 ,  C12N2310/3519 ,  C12N2320/10

摘要： A regulatable gene expression construct comprising a nucleic acid molecule comprising a two-way riboswitch operably linked to a target sequence. Also provided is a library screening strategy for efficient creation of target-specific riboswitches. A theophylline-repressible and IPTG-inducible riboswitch device achieves portable control of gene expression control in a ‘two-way’ manner. The default state of target genes is ON; the targets are switched off by adding theophylline, and switched back to the ON-state by adding IPTG without changing growth medium. The riboswitch device regulates gene expression in a portable, adjustable, and two-way manner with a variety of scientific and biotechnological applications.

摘要翻译： 一种可调节基因表达构建体，其包含核酸分子，其包含可操作地连接到靶序列的双向核糖开关。 还提供了一种用于有效创建目标特异性核糖开关的图书馆筛选策略。 茶碱抑制性和IPTG诱导型核糖开关器件以双向方式实现基因表达控制的便携式控制。 目标基因的默认状态为ON; 通过添加茶碱来关闭目标，并通过添加IPTG而不改变生长培养基而切换回ON状态。 核糖开关器件通过各种科学和生物技术应用以便携式，可调节和双向的方式调节基因表达。

公开(公告)号：US07781190B2

公开(公告)日：2010-08-24

申请号：US10892579

申请日：2004-07-16

申请人： Jian-Dong Huang ,  Xin-Mei Zhang ,  Julian Alexander Tanner

发明人： Jian-Dong Huang ,  Xin-Mei Zhang ,  Julian Alexander Tanner

IPC分类号： C12N15/64 ,  C12N15/70

CPC分类号： C12N15/10

摘要： This invention provides a method for combining overlapping DNA molecules comprising: (a) providing first and second DNA fragments, the first having a region homologous to a region in the second; (b) tagging the first DNA fragment with a selectable marker; (c) cloning the first DNA sequence into a retrieval vector to form a DNA-vector complex; (d) linearizing the DNA-vector complex; and (e) inserting the first DNA fragment from the DNA-vector complex into the second DNA fragment using homologous recombination to form a combined DNA molecule; and (f) removing the selectable marker, thereby generating a combined DNA molecule. The invention further provides a vector for retrieving and inserting a selected DNA molecule into a target DNA molecule.

摘要翻译： 本发明提供了一种组合重叠DNA分子的方法，包括：（a）提供第一和第二DNA片段，第一DNA片段具有与第二DNA片段中的区域同源的区域; （b）用选择标记标记第一个DNA片段; （c）将第一DNA序列克隆到检索载体中以形成DNA载体复合物; （d）使DNA-载体复合物线性化; 和（e）使用同源重组将来自DNA-载体复合物的第一个DNA片段插入第二个DNA片段以形成组合的DNA分子; 和（f）除去选择性标记，从而产生组合的DNA分子。 本发明还提供了用于检索并将选定的DNA分子插入靶DNA分子的载体。

公开(公告)号：US20050084970A1

公开(公告)日：2005-04-21

申请号：US10892579

申请日：2004-07-16

申请人： Jian-Dong Huang ,  Xin-Mei Zhang ,  Julian Tanner

发明人： Jian-Dong Huang ,  Xin-Mei Zhang ,  Julian Tanner

IPC分类号： C12N15/10 ,  C12N15/867 ,  C12P19/34

CPC分类号： C12N15/10

摘要： This invention provides a method for combining overlapping DNA molecules comprising: (a) providing first and second DNA fragments, the first having a region homologous to a region in the second; (b) tagging the first DNA fragment with a selectable marker; (c) cloning the first DNA sequence into a retrieval vector to form a DNA-vector complex; (d) linearizing the DNA-vector complex; and (e) inserting the first DNA fragment from the DNA-vector complex into the second DNA fragment using homologous recombination to form a combined DNA molecule; and (f) removing the selectable marker, thereby generating a combined DNA molecule. The invention further provides a vector for retrieving and inserting a selected DNA molecule into a target DNA molecule.

摘要翻译： 本发明提供了一种组合重叠DNA分子的方法，包括：（a）提供第一和第二DNA片段，第一DNA片段具有与第二DNA片段中的区域同源的区域; （b）用选择标记标记第一个DNA片段; （c）将第一DNA序列克隆到检索载体中以形成DNA载体复合物; （d）使DNA-载体复合物线性化; 和（e）使用同源重组将来自DNA-载体复合物的第一个DNA片段插入第二个DNA片段以形成组合的DNA分子; 和（f）除去选择性标记，从而产生组合的DNA分子。 本发明还提供了用于检索并将选定的DNA分子插入靶DNA分子的载体。