CLONING AND EXPRESSION OF FULL LENGTH HUMAN ORFS

全长人类 ORFS 的克隆和表达

• 批准号: 6173776
• 负责人: JOHN J CARRINO
• 金额: $ 39.36万
• 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/6173776
• 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 将基因组学与功能基因组学联系起来的技术。 它们共同使科学家能够整合来自 将基因组分解为蛋白质，即生命的功能单位。 英杰公司的 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元 硬件。 我们最近开发了一种自动克隆和 表达系统围绕我们的多项专利和专有技术构建 许可证。 我们将使用该系统将全长人类 cDNA 克隆到 哺乳动物表达载体，可进行功能表征 基因产物。 申请和研究计划将分开 分为 3 个不同的重点领域。 生成的全长人类 cDNA 这些重点领域采用的每项策略都将被克隆 转化为表达质粒，测序并在 CHO 细胞中表达 验证完整性。 该研究计划旨在重点关注全面 现有数据库中的长度序列来指导模板组装和 基于 RT-PCR 的全长模板获取策略作为第一个 水平策略，然后利用现有的 EST 同源性标准 优先获取癌症相关的未知 5' 末端 cDNA 拟议的商业应用： 克隆的癌症相关基因的营销将涉及以下策略： 迅速将第一个也是最大的全系列产品推向市场 表达（和其他）载体中癌症相关基因的长度。营销 这些基因将包括 Invitrogen 促销的全部范围 策略例如： 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. 通过互联网访问可搜索的克隆基因数据库 4.向行业内的目标受众定向发送促销邮件 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