EXON TRAPPING - A GENETIC SCREEN TO IDENTIFY CANDIDATE TRANSCRIBED SEQUENCES IN CLONED MAMMALIAN GENOMIC DNA

被引:163
作者
DUYK, GM
KIM, SW
MYERS, RM
COX, DR
机构
[1] UNIV CALIF SAN FRANCISCO,DEPT PHYSIOL,SAN FRANCISCO,CA 94143
[2] UNIV CALIF SAN FRANCISCO,DEPT BIOCHEM & BIOPHYS,SAN FRANCISCO,CA 94143
[3] UNIV CALIF SAN FRANCISCO,DEPT PSYCHIAT,SAN FRANCISCO,CA 94143
关键词
human genetics; retroviral vectors; RNA splicing;
D O I
10.1073/pnas.87.22.8995
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Identification and recovery of transcribed sequences from cloned mammalian genomic DNA remains an important problem in isolating genes on the basis of their chromosomal location. We have developed a strategy that facilitates the recovery of exons from random pieces of cloned genomic DNA. The basis of this 'exon trapping' strategy is that, during a retroviral life cycle, genomic sequences of nonviral origin are correctly spliced and may be recovered as a cDNA copy of the introduced segment. By using this genetic assay for cis-acting sequences required for RNA splicing, we have screened ≃20 kilobase pairs of cloned genomic DNA and have recovered all four predicted exons.
引用
收藏
页码:8995 / 8999
页数:5
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