公开(公告)号：US07020560B2

公开(公告)日：2006-03-28

申请号：US09949015

申请日：2001-09-06

Applicant: Gary S. Sayler ,  James T. Fleming ,  Bruce Applegate ,  Michael L. Simpson

Inventor： Gary S. Sayler ,  James T. Fleming ,  Bruce Applegate ,  Michael L. Simpson

IPC: G01N21/00 ,  G01N27/00 ,  G01N33/48

CPC classification number: C12Q1/6897 ,  B82Y10/00 ,  G01N33/5005 ,  G01N33/5438 ,  G06N3/002

Abstract: A method for cell-based combinatorial logic includes the steps of providing at least one genetically engineered cell, the genetically engineered cell having at least one transcriptional unit. The transcriptional unit includes a gene and a promoter, wherein application of a stimulus to the promoter results in the expression of a gene product. An energetic or chemical stimulus is applied to activate the promoter, wherein the detection of an output signal corresponds to the presence of a gene product. The cell can include a plurality of transcriptional units configured to form logic gates. The logic gates of a plurality of cells can be operably interconnected by release of output signals, such as chemical stimuli.

Abstract translation: 一种用于基于小区的组合逻辑的方法包括以下步骤：提供至少一个经遗传工程化的细胞，所述基因工程细胞具有至少一个转录单位。 转录单位包括基因和启动子，其中向启动子施加刺激物导致基因产物的表达。 施加能量或化学刺激以激活启动子，其中输出信号的检测对应于基因产物的存在。 该小区可以包括被配置成形成逻辑门的多个转录单元。 多个单元的逻辑门可以通过释放诸如化学刺激的输出信号来可操作地互连。

公开(公告)号：US06090593A

公开(公告)日：2000-07-18

申请号：US78283

申请日：1998-05-13

Applicant: James T. Fleming ,  Gary S. Sayler

Inventor： James T. Fleming ,  Gary S. Sayler

IPC: C12Q1/68 ,  C12P19/34 ,  C07H21/04

CPC classification number: C12Q1/6809 ,  C12Q1/6888 ,  C12Q1/689

Abstract: The differential display (DD) technique, widely used previously for eukaryotic gene discovery, is optimized to detect differential mRNA transcription (an expressed gene) from both pure culture and soil derived bacterial RNA. A model system using toluene induction of TodC1 in Pseudomonas putida F1 is used to optimize the procedure. Once optimized, an arbitrary primer for the RT step in conjunction with the same arbitrary primer and a Shine-Dalgarno (SD) primer for the PCR reaction is used to detect the expressed gene. The invention thus provides a method for discovery and acquisition of novel genes from environmental microbial communities that avoids the traditional steps and inherent bias due to the culturing of environmental isolates.

Abstract translation: 针对真核基因发现广泛使用的差异显示（DD）技术进行了优化，以检测来自纯培养物和土壤细菌RNA的差异mRNA转录（表达基因）。 在恶臭假单胞菌F1中使用TodC1甲苯诱导的模型系统用于优化程序。 一旦优化，使用RT步骤的任意引物连同相同的任意引物和用于PCR反应的Shine-Dalgarno（SD）引物来检测表达的基因。 因此，本发明提供了一种从环境微生物群落中发现和获取新基因的方法，其避免了由于环境分离物的培养而导致的传统步骤和固有偏差。