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

被引:164
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
相关论文
共 28 条
[1]   SEARCHING FOR CODING SEQUENCES IN THE MAMMALIAN GENOME - THE H-2K REGION OF THE MOUSE MHC IS REPLETE WITH GENES EXPRESSED IN EMBRYOS [J].
ABE, K ;
WEI, JF ;
WEI, FS ;
HSU, YC ;
UEHARA, H ;
ARTZT, K ;
BENNETT, D .
EMBO JOURNAL, 1988, 7 (11) :3441-3449
[2]   TRANSGENES AS PROBES FOR ACTIVE CHROMOSOMAL DOMAINS IN MOUSE DEVELOPMENT [J].
ALLEN, ND ;
CRAN, DG ;
BARTON, SC ;
HETTLE, S ;
REIK, W ;
SURANI, MA .
NATURE, 1988, 333 (6176) :852-855
[3]   CHROMOSOME CRAWLING IN THE MHC [J].
BELL, J .
TRENDS IN GENETICS, 1989, 5 (09) :289-290
[4]   CPG ISLANDS AS GENE MARKERS IN THE VERTEBRATE NUCLEUS [J].
BIRD, AP .
TRENDS IN GENETICS, 1987, 3 (12) :342-347
[5]  
BROWN AMC, 1987, DNA CLONING, V3, P189
[6]   CONSTRUCTION AND APPLICATIONS OF A HIGHLY TRANSMISSIBLE MURINE RETROVIRUS SHUTTLE VECTOR [J].
CEPKO, CL ;
ROBERTS, BE ;
MULLIGAN, RC .
CELL, 1984, 37 (03) :1053-1062
[7]   USE OF RETROVIRAL VECTORS FOR MAPPING OF SPLICE SITES IN COTTONTAIL RABBIT PAPILLOMAVIRUS [J].
DOSTATNI, N ;
YANIV, M ;
DANOS, O ;
MULLIGAN, RC .
JOURNAL OF GENERAL VIROLOGY, 1988, 69 :3093-3100
[8]  
DOUGHERTY J, 1986, MOL CELL BIOL, V63, P4387
[9]   LIPOFECTION - A HIGHLY EFFICIENT, LIPID-MEDIATED DNA-TRANSFECTION PROCEDURE [J].
FELGNER, PL ;
GADEK, TR ;
HOLM, M ;
ROMAN, R ;
CHAN, HW ;
WENZ, M ;
NORTHROP, JP ;
RINGOLD, GM ;
DANIELSEN, M .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (21) :7413-7417
[10]   SV40-TRANSFORMED SIMIAN CELLS SUPPORT THE REPLICATION OF EARLY SV40 MUTANTS [J].
GLUZMAN, Y .
CELL, 1981, 23 (01) :175-182