公开(公告)号：US20060160222A1

公开(公告)日：2006-07-20

申请号：US11006470

申请日：2004-12-06

申请人： Kevin Rozwadowski ,  Derek Lydiate

发明人： Kevin Rozwadowski ,  Derek Lydiate

IPC分类号： C12N15/82

CPC分类号： A01H1/02 ,  C12N15/102 ,  C12N15/8213

摘要： The present invention provides methods of modifying in vivo mutagenesis or homeologous recombination in a eukaryote. The method of modifying in vivo mutagenesis involves transforming a eukaryote with a nucleotide sequence capable of expressing a wild-type prokaryotic MutS, MutL, MutH, MutU, NLS-MutS, NLS-MutL, NLS-MutH, NLS-MutU protein, or a combination thereof, and expressing the protein. A method of modifying recombination between homeologous chromosomes in an allopolyploid eukaryotic organism comprising, expressing a nucleotide sequence encoding prokaryotic NLS-MutS in combination with one or more than one of NLS-MutL, NLS-MutH, NLS-MutU, within a germ cell of the allopolyploid eukaryotic organism is also disclosed.

摘要翻译： 本发明提供在真核生物体内改变体内诱变或同源重组的方法。 修饰体内诱变的方法包括用能够表达野生型原核MutS，MutL，MutH，MutU，NLS-MutS，NLS-MutL，NLS-MutH，NLS-MutU蛋白或NLS-MutU蛋白的核苷酸序列转化真核生物 并组合，并表达蛋白质。 一种修饰异源多倍体真核生物中的同源染色体之间的重组的方法，其包括将编码原核NLS-MutS的核苷酸序列与NLS-MutL，NLS-MutH，NLS-MutU中的一种或多种N 还披露了异倍体多倍体真核生物。

公开(公告)号：US20050250207A1

公开(公告)日：2005-11-10

申请号：US10516779

申请日：2003-06-05

申请人： Kevin Rozwadowski ,  Derek Lydiate

发明人： Kevin Rozwadowski ,  Derek Lydiate

IPC分类号： A61K48/00 ,  C12N1/18 ,  C12N5/04 ,  C12N5/06 ,  C12N9/12 ,  C12N15/10 ,  C12N15/74 ,  C12N15/85 ,  C12N15/90

CPC分类号： C12N15/102 ,  A01K2217/05 ,  A61K48/00 ,  C12N9/1276 ,  C12N15/902

摘要： The invention provides methods and nucleic acid constructs that may be used to modify a nucleic acid of interest at a target locus within the genome of a host. In some aspects, the invention contemplates producing in vivo a gene targeting substrate (GTS), which may be comprised of both DNA and RNA components. The gene targeting substrate may comprise a gene targeting nucleotide sequence (GTNS), which is homologous to the target locus, but comprises a sequence modification compared to the target locus. The gene targeting substrate may be produced by reverse transcription of a gene targeting message RNA (gtmRNA). The gene targeting message RNA may be folded for self-priming for reverse transcription by a reverse transcriptase. The gene targeting message RNA may in turn be the product of transcription of a gene targeting construct (GTC) encoding the gene targeting message RNA. The gene targeting construct may for example be a DNA sequence integrated into the genome of the host, or integrated into an extrachromosomal element. Following expression of the gene targeting systems of the invention, hosts may for example be selected having genomic modifications at a target locus that correspond to the sequence modification present on the gene targeting nucleotide sequence. In some embodiments, the structure of retrons may be adapted for use in the gene targeting systems of the invention.

摘要翻译： 本发明提供可用于在宿主基因组内的靶基因座处修饰目标核酸的方法和核酸构建体。 在一些方面，本发明考虑在体内产生靶基质（GTS），其可以由DNA和RNA组分两者组成。 基因靶向底物可以包含与靶基因座同源的基因靶向核苷酸序列（GTNS），但与目标基因座相比包含序列修饰。 基因靶向底物可以通过靶向消息RNA（gtmRNA）的基因的逆转录产生。 基因靶向消息RNA可以被折叠用于通过逆转录酶进行逆激酶的自引发。 基因靶向消息RNA可能又是编码基因靶向消息RNA的基因靶向构建体（GTC）的转录产物。 基因靶向构建体可以例如是整合到宿主基因组中的DNA序列，或整合到染色体外元件中。 在表达本发明的基因靶向系统之后，例如可以选择宿主在靶基因座具有与基因靶向核苷酸序列上存在的序列修饰相对应的基因组修饰。 在一些实施方案中，维管束的结构可适用于本发明的基因靶向系统。

公开(公告)号：US20060123506A1

公开(公告)日：2006-06-08

申请号：US10516753

申请日：2003-06-06

申请人： Abdelali Hannoufa ,  Derek Lydiate ,  Ming-Jun Gao

发明人： Abdelali Hannoufa ,  Derek Lydiate ,  Ming-Jun Gao

IPC分类号： A01H1/00 ,  C12N15/82

CPC分类号： C07K14/415 ,  C12N15/63 ,  C12N15/8216 ,  C12N15/8217

摘要： The invention provides a method to regulate expression of a gene of interest in a plant comprising, introducing into the plant a first nucleotide sequence comprising, the gene of interest operatively linked to a first regulatory region, and an operator sequence capable of binding a fusion protein, and a second nucleotide sequence comprising a second regulatory region in operative association with a nucleotide sequence encoding the fusion protein. The fusion protein comprising, a DNA binding protein, or a portion thereof, capable of binding the operator sequence, and a recruitment factor protein, or a portion thereof, capable of binding a chromatin remodelling protein. In this manner, expression of the second nucleotide sequence produces the fusion protein that regulates expression of the gene of interest.

公开(公告)号：US20050278809A1

公开(公告)日：2005-12-15

申请号：US11067425

申请日：2005-02-22

申请人： Abdelali Hannoufa ,  Derek Lydiate ,  Ming-Jun Gao

发明人： Abdelali Hannoufa ,  Derek Lydiate ,  Ming-Jun Gao

IPC分类号： A01H1/00 ,  C07H21/04 ,  C12N5/04 ,  C12N15/82

CPC分类号： C12N15/8217

摘要： The invention provides a method to regulate expression of a gene of interest in a plant comprising, introducing into the plant a first nucleotide sequence comprising, the gene of interest operatively linked to a first regulatory region, and an operator sequence capable of binding a fusion protein, and a second nucleotide sequence comprising a second regulatory region in operative association with a nucleotide sequence encoding the fusion protein. The fusion protein comprising, a DNA binding protein, or a portion thereof, capable of binding the operator sequence, and a recruitment factor protein, or a portion thereof, capable of binding a chromatin remodelling protein. In this manner, expression of the second nucleotide sequence produces the fusion protein that regulates expression of the gene of interest.

摘要翻译： 本发明提供了调节植物中目的基因表达的方法，包括：向植物中引入第一核苷酸序列，其包含可操作地连接到第一调节区的目的基因和能够结合融合蛋白的操纵子序列 和包含与编码融合蛋白的核苷酸序列有效结合的第二调节区的第二核苷酸序列。 包含能够结合操作者序列的DNA结合蛋白或其部分的融合蛋白，以及能够结合染色质重塑蛋白的募集因子蛋白或其部分。 以这种方式，第二核苷酸序列的表达产生调节感兴趣的基因表达的融合蛋白。