基因组隔离子的高通量鉴定方法

侯春晖; 牛龙见; 张越东; 何娜; 孙加雷; 黄盈长

侯春晖

南方科技大学

南方科技大学

518055 广东省深圳市南山区西丽学苑大道1088号

南方科技大学

518055 广东省深圳市南山区西丽学苑大道1088号 (广东,深圳,南山区)

南方科技大学

518055 广东省深圳市南山区西丽学苑大道1088号 (广东,深圳,南山区)

CN201710659482.5

2017-08-04

CN107604043B

2019-04-16

2019-04-16

未缴年费

2024-06-25

授权发明

第一

本发明公开了基因组隔离子的高通量鉴定方法，包括：1)构建报告载体：报告载体依次具有启动子、ORF区、内含子和隔离子候选序列插入区和增强子；增强子用于增强上游序列的表达，隔离子候选序列插入区插入有隔离子候选序列；2)将报告载体表达转录；3)检测转录序列的编辑后转录情况，确定隔离子候选序列是否为隔离子。该方法构建的报告载体库可以覆盖90％到99％的基因组非编码区，不需要蛋白位置信息，全部分析；将所构建的库转入细胞内，可以同时检测几乎整个基因组序列是否具有隔离子活性，可以大规模并行分析，避免单个、少量的分析；因为所构建库覆盖几乎所有位点，因此未知区域同样可以被检测分析。

The invention discloses a high-throughput identification method for a genome insulator. The high-throughput identification method for the genome insulator includes the steps of (1) building a reportervector, specifically, the reporter vector sequentially comprises a promoter, an ORF region, an intron, an insulator candidate sequence insertion region and an enhancer, the enhancer is used for enhancing expression of an upstream sequence, and an insulator candidate sequence is inserted in the insulator candidate sequence insertion region; (2) expressing and transcribing the reporter vector; and(3) detecting the transcribing condition of the edited transcribing sequence, and determining whether the insulator candidate sequence is the insulator or not. A reporter vector library built by the method can cover 90-99% of genome noncoding regions, protein position information is not needed, and overall analysis is achieved; the built library is transferred into a cell, whether nearly the wholegenomic sequence has insulator activity or not can be detected at the same time, large-scale parallel analysis can be carried out, and single and small-quantity analysis is avoided; and since the built library covers nearly all sites, zones of ignorance can be also detected and analyzed.

C12Q1/68

（+PATENT GRANT）[2019-04-16][CN]

1

1

唐致明

广州嘉权专利商标事务所有限公司

PatSnap

侯春晖,牛龙见,张越东,等. 基因组隔离子的高通量鉴定方法[P]. 2019-04-16.