公开(公告)号：US20220119823A1

公开(公告)日：2022-04-21

申请号：US17453238

申请日：2021-11-02

申请人： National University of Singapore ,  Massachusetts Institute of Technology

发明人： Heng Phon Too ,  Ruiyang Zou ,  Gregory N. Stephanopoulos

IPC分类号： C12N15/67 ,  C12N15/63 ,  C07H21/00 ,  C12N15/69 ,  C12N15/52 ,  C12N15/68

摘要： The invention provides, inter alia, a nucleic acid (e.g. expression vector) that comprises at least a first coding sequence and a second coding sequence. Each conding sequence is under the control of an inducible promoter of defined strength. Different promoters can have different strengths. Each promoter is responsive to the same inducer. The invention also provides: methods of expressing coding regions, methods of making a product of a multi-enzyme pathway, and methods of optimizing the yield of a product of a multi-enzyme metabolic pathway using the nucleic acids provided by the invention. Also disclosed is a method of non-enzymatic gene cloning useful for practicing the invention.

公开(公告)号：US11149279B2

公开(公告)日：2021-10-19

申请号：US16636713

申请日：2018-03-26

申请人： CELL AND GENE THERAPY LTD ,  OBSCHESTVO S OGRANICHENNOI OTVETSTVENNOSTJU “PRORYVNYE INNOVATSIONNYE TEKHNOLOGII”

发明人： Natalia Savelieva

IPC分类号： C12N15/69 ,  C12N9/14 ,  C12N15/70 ,  C12R1/19

摘要： Disclosed is a gene therapy DNA vector VTvaf17 for genetic modification of animal and human cells having SEQ ID No. 1, and methods for its synthesis, involving constructing vectors containing a promoter region of human elongation factor EF1A, a polylinker with sites for restriction endonucleases, a transcription terminator, a polyadenylation sequence of human growth factor, a regulatory element of transposon Tn10 allowing for antibiotic-free positive selection, an origin of replication, and a kanamycin resistance gene. Escherichia coli strain SCS110-AF is also provided by the present invention. The method for creating the strain involves constructing a linear DNA fragment containing regulatory element of transposon Tn10, a levansucrase gene, sacB, a chloramphenicol resistance gene, and two homologous sequences. The E. coli cells are transformed by electroporation and clones surviving chloramphenicol are chosen. The invention further discloses Escherichia coli strain SCS110-AF/VTvaf17, which carries DNA vector VTvaf17, and methods for its synthesis.

公开(公告)号：US20210269508A1

公开(公告)日：2021-09-02

申请号：US17320661

申请日：2021-05-14

申请人： Cellastra Inc.

发明人： Bradley G. THOMPSON

IPC分类号： C07K14/79 ,  A61K31/198 ,  A61P17/02 ,  A61K48/00 ,  C12N15/69

摘要： The present disclosure relates to the composition of one or more agents, therapies, treatments, and methods of use of the agents and/or therapies and/or treatments for upregulating production of lactoferrin or a sub-peptide of lactoferrin. Embodiments of the present disclosure can be used as a therapy or a treatment of adhesions or scarring.

公开(公告)号：US20200347419A1

公开(公告)日：2020-11-05

申请号：US16760662

申请日：2018-05-22

申请人： Ningxia Eppen Biotech Co., Ltd ,  Institute of Microbiology, Chinese Academy of Sciences

发明人： Tingyi WEN ,  Chen ZHANG ,  Xiuling SHANG ,  Xin CHAI ,  Yun ZHANG ,  Shuwen LIU ,  Guoqiang WANG ,  Zhongcai LI

IPC分类号： C12P13/08 ,  C12N15/64 ,  C12N15/69

摘要： A recombinant bacterium for producing L-lysine, a construction method thereof, and a method for producing L-lysine by using the recombinant bacterium. The recombinant bacterium has increased expression and/or activity of asparaginase compared to a starting bacterium.

公开(公告)号：US20190264216A1

公开(公告)日：2019-08-29

申请号：US16409586

申请日：2019-05-10

申请人： Intact Genomics, Inc.

发明人： Chengcang Charles Wu

IPC分类号： C12N15/80 ,  C12N15/69 ,  C12N15/52 ,  C12N15/10

摘要： Fungal artificial chromosome (FAC) vectors are disclosed. A vector can be replicated in a bacterial or a fungal host, and can comprise an insert of heterologous DNA up to about 500 kilobases. A vector can be used for cloning and expressing a secondary metabolite (SM) gene cluster. An insert sequence can be modified by homologous recombination. A vector can be a plasmid comprising bacterial and fungal origins of replication, as well as bacterial and fungal selection marker genes. Also disclosed are vectors that can be integrated into a fungal genome, and dual function vectors which can be replicated in a bacterial or a fungal host and can also be integrated into a fungal genome. Also disclosed are methods of generating plasmid libraries including vectors comprising intact SM gene clusters.

公开(公告)号：US10131913B2

公开(公告)日：2018-11-20

申请号：US14681351

申请日：2015-04-08

申请人： CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE (CNRS)

发明人： Marcel Mechali ,  Philippe Coulombe ,  Christelle Cayrou ,  Eric Rivals ,  Paulina Prorok

IPC分类号： C12N15/64 ,  C12N15/69

摘要： A method for initiating the replication of a deoxyribonucleic acid molecule includes inserting into the DNA at least one nucleic acid molecule representing a multicellular DNA replication origin. The replication origin contains at least nine nucleotides and contains at least three uninterrupted origin repeating elements (ORE), each ORE having the sequence N3GN4, wherein N3 is T or G and N4 is G or C. The method can confer autonomous replication properties to a non-self-replicating DNA molecule. A process for preparing a vector for use in said methods is also presented.

公开(公告)号：US10076564B2

公开(公告)日：2018-09-18

申请号：US14787098

申请日：2014-04-25

申请人： KATHOLIEKE UNIVERSITEIT LEUVEN

发明人： Kai Dallmeier ,  Johan Neyts

IPC分类号： A61K39/12 ,  C12N7/00 ,  C12N15/69 ,  A61K39/00

CPC分类号： A61K39/12 ,  A61K2039/5254 ,  C12N7/00 ,  C12N15/69 ,  C12N2770/24134

摘要： The invention relates to the use of a bacterial artificial chromosome (BAC) for the preparation of a vaccine, wherein the BAC comprises an inducible bacterial ori sequence for amplification of the BAC to more than 10 copies per bacterial cell. Plus a viral expression cassette comprising a cDNA of an attenuated RNA virus genome and comprising cis-regulatory elements for transcription of said viral cDNA in mammalian cells and for processing of the transcribed RNA into infectious viral RNA.

公开(公告)号：US09670495B2

公开(公告)日：2017-06-06

申请号：US14770074

申请日：2014-02-25

申请人： UNIVERSITY OF WASHINGTON THROUGH ITS CENTER FOR COMMERCIALIZATION

发明人： Maitreya Dunham ,  Ivan Liachko

IPC分类号： C12Q1/68 ,  C12Q1/00 ,  C12N15/81 ,  C12N15/69

CPC分类号： C12N15/81 ,  C12N15/69 ,  C12N15/815

摘要： A DNA sequence that functions as an origin in many different yeast species. From 1 to 17 mutations can be introduced into this sequence to improve its function across multiple yeasts. The resulting synthetic DNA sequence confers stable plasmid replication function in all yeast species tested, including but not limited to Saccharomyces cerevisiae, Lachancea kluyveri, Kluyveromyces lactis, Kluyveromyces wickerhammii, Hansenula polymorpha, and Pichia pastoris. Also provided are sequences that function as an optimal origin in the industrially useful Pichia pastoris.

公开(公告)号：US6022716A

公开(公告)日：2000-02-08

申请号：US058746

申请日：1998-04-10

申请人： Ilya Chumakov ,  Hiroaki Tanaka

发明人： Ilya Chumakov ,  Hiroaki Tanaka

IPC分类号： C12N15/09 ,  C12N9/38 ,  C12N15/10 ,  C12N15/63 ,  C12N15/69 ,  C12Q1/68 ,  C12N15/11 ,  C12N15/64 ,  C12Q1/00 ,  C12Q1/04

CPC分类号： C12Q1/6841 ,  C12N15/102 ,  C12N15/63 ,  C12N15/69 ,  C12N9/16 ,  C12N9/2471 ,  C12Q1/6869 ,  C12Y302/01023

摘要： High throughput DNA sequencing vectors for generating nested deletions using enzymatic techniques and/or transposition-based techniques are disclosed. Methods of constructing contigs of long DNA sequences and methods of generating nested deletions are also disclosed. A truncated lacZ derivative useful in measuring the copy number of the lacZ derivative in a host cell is also disclosed.

公开(公告)号：US5702916A

公开(公告)日：1997-12-30

申请号：US449958

申请日：1995-05-25

申请人： S.o slashed.ren Molin ,  Poul Kirketerp Andersson ,  Kenn Axo Gerdes ,  Per Klemm

发明人： S.o slashed.ren Molin ,  Poul Kirketerp Andersson ,  Kenn Axo Gerdes ,  Per Klemm

IPC分类号： A61K39/00 ,  C12N15/00 ,  C12N15/68 ,  C12N15/69 ,  C12P21/00 ,  C12N1/21 ,  C12N15/63

CPC分类号： C12N15/69 ,  C12N15/00 ,  C12N15/68 ,  A61K39/00

摘要： A replicon, in which a nucleotide sequence encoding a cell killing function is regulatably expressed when the replicon is harbored in one type of host cell (primary host cell), so that cells harboring the replicon are killed under conditions under which the cell killing function is expressed, and the nucleotide sequence encoding the cell killing function is regulatably or constitutively expressed when the replicon is harbored in another type of host cell (secondary host cell), so that cells harboring the replicon are invariably killed or killed under conditions under which the cell killing function is expressed, may be used in a method of active biological containment of cells under defined environmental conditions. The biological containment principle may be utilized in the industrial production of a biosynthetic product by recombinant DNA techniques, when deliberately releasing a genetically engineered microorganism to the natural environment or in the preparation of a live vaccine. The expression of the cell killing function may be regulated by means of a promoter.

摘要翻译： 当复制子存在于一种类型的宿主细胞（主要宿主细胞）中时，可复制表达编码细胞杀伤功能的核苷酸序列的复制子，使得携带复制子的细胞在细胞杀伤功能为 表达，并且当复制子宿主在另一种类型的宿主细胞（次级宿主细胞）中时，编码细胞杀伤功能的核苷酸序列是可调节的或组成型表达的，使得携带复制子的细胞在细胞的条件下总是被杀死或杀死 表达杀伤功能，可用于在确定的环境条件下对细胞的活性生物遏制的方法。 当将遗传工程微生物故意释放到天然环境或活疫苗的制备中时，可通过重组DNA技术在生物合成产物的工业生产中利用生物遏制原理。 细胞杀伤功能的表达可以通过启动子调节。