CLONING AND EXPRESSION OF FULL LENGTH HUMAN ORFS

全长人类 ORFS 的克隆和表达

• 批准号: 6210224
• 负责人: JOHN A HEYMAN
• 金额: $ 1.03万
• 依托单位: INVITROGEN CORPORATION
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6210224
• 关键词: CHO cells; complementary DNA; gene expression; human genetic material tag; molecular cloning; nucleic acid sequence; plasmids; polymerase chain reaction; tissue /cell culture; transfection /expression vector

DESCRIPTION: (Applicant's Description) Gene cloning and expression are enabling technologies linking genomics to functional genomics. Together, they allow scientists to convent the genetic information from a genome into proteins, the functional units of life. Invitrogen's cloning and expression vectors are similar to an operating system for conversion of genetic information into functional units via cellular hardware. We have recently developed an automated cloning and expression system built around several of our patents and exclusive licenses. We will use this system to clone full-length human cDNAs into a mammalian expression vector to allow functional characterization of the gene products. The application and research plan will be divided into 3 separate areas of focus. The full-length human cDNAs generated by each of the strategies employed in these focus areas will be cloned into expression plasmids, sequenced, and expressed in CHO cells to verify integrity. The research plan is designed to focus on the full- length sequences in existing databases to guide template assembly and RT-PCR-based strategies of full-length template acquisition as a first- level strategy, and then utilize the existing EST homology criteria to prioritize the acquisition of the unknown 5' ends of cancer-related CDNAs PROPOSED COMMERCIAL APPLICATION: Marketing of the cloned cancer-related genes will involve a strategy of quickly bringing to market the first and largest collection the full- length cancer-related genes in expression (and other) vectors. Marketing of these genes will include the full range of Invitrogen's promotional tactics such as: 1. Advertisements in scientific journals and publications 2. Publication of articles in Invitrogen's newsletter Expressions (6 issues annually) 3. Internet access to a searchable database of cloned genes 4. Directed promotional mailings to target audiences in industry and academia 5. Public relations and press releases 6. Trade shows 7. Scientific seminars, and workshops

描述：（申请人的描述）基因克隆和表达是 使技术将基因组学与功能基因组学联系起来。 它们共同使科学家能够从修道院获取遗传信息 将基因组转化为蛋白质，生命的功能单位。 Invitrogen的 克隆和表达载体类似于操作系统 通过细胞将遗传信息转化为功能单位 硬件. 我们最近开发了一种自动克隆技术， 表达系统围绕我们的几项专利和独家 许可证 我们将用这个系统克隆全长人类cDNA到 哺乳动物表达载体，以允许功能表征 基因产物。 申请和研究计划将分为 分为三个不同的重点领域。 产生的全长人类cDNA 在这些重点领域采用的每一种战略将被克隆 转化到表达质粒中，测序并在CHO细胞中表达， 验证完整性。 该研究计划的目的是集中在完整的- 现有数据库中的长度序列来指导模板组装， 基于RT-PCR的全长模板获取策略作为第一步， 水平的策略，然后利用现有的EST同源性标准， 优先获得癌症相关的未知的5'末端， 的cdna 拟定商业应用： 克隆的癌症相关基因的市场营销将涉及以下策略： 迅速将第一个也是最大的系列推向市场， 长度癌症相关基因的表达（和其他）载体。营销 这些基因将包括英杰公司的全方位的宣传， 战术，例如： 1.科学期刊和出版物上的广告 2.在Invitrogen的时事通讯Expressions上发表文章 （每年6期） 3.通过因特网访问可检索的克隆基因数据库 4.向工业界的目标受众发送促销邮件 和学术界 5.公共关系和新闻稿 6.贸易展览 7.科学研讨会和讲习班

项目成果

Genome-scale cloning and expression of individual open reading frames using topoisomerase I-mediated ligation.

使用拓扑异构酶 I 介导的连接对单个开放阅读框进行基因组规模克隆和表达。

• 发表时间: 1999
• 期刊: Genome research
• 影响因子: 7
• 作者: Heyman,JA;Cornthwaite,J;Foncerrada,L;Gilmore,JR;Gontang,E;Hartman,KJ;Hernandez,CL;Hood,R;Hull,HM;Lee,WY;Marcil,R;Marsh,EJ;Mudd,KM;Patino,MJ;Purcell,TJ;Rowland,JJ;Sindici,ML;Hoeffler,JP
• 通讯作者: Hoeffler,JP

High-throughput expression of fusion proteins.

融合蛋白的高通量表达。

• DOI: 10.1016/s0076-6879(00)28416-4
• 发表时间: 2000
• 期刊: Methods in enzymology
• 作者: Nasoff,M;Bergseid,M;Hoeffler,JP;Heyman,JA
• 通讯作者: Heyman,JA