Identification of fission yeast nuclear markers using random polypeptide fusions with green fluorescent protein

被引:72
作者
Sawin, KE [1 ]
Nurse, P [1 ]
机构
[1] IMPERIAL CANC RES FUND, CELL CYCLE LAB, LONDON WC2A 3PX, ENGLAND
关键词
D O I
10.1073/pnas.93.26.15146
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
We describe a method for identifying genes encoding proteins with stereospecific intracellular localizations in the fission yeast Schizosaccharomyces pombe. Yeast are transformed with a gene library in which S, pombe genomic sequences are fused to the gene encoding the Aequorea victoria green fluorescent protein (GFP), and intracellular localizations are subsequently identified by rapid fluorescence screening in vivo. In it model application of these methods to the fission yeast nucleus, we have identified several novel genes whose products are found in specific nuclear regions, including chromatin, the nucleolus, and the mitotic spindle, and sequence similarities between some of these genes and previously identified genes encoding nuclear proteins have validated the approach, These methods will he useful in identifying additional components of the S, pombe nucleus, and further extensions of this approach should also be applicable to a more comprehensive identification of the elements of intracellular architecture in fission yeast.
引用
收藏
页码:15146 / 15151
页数:6
相关论文
共 30 条
[1]  
AASLAND R, 1995, NUCLEIC ACIDS RES, V23, P3168
[2]   EXPRESSION AND ANALYSIS OF THE GREEN FLUORESCENT PROTEIN GENE IN THE FISSION YEAST SCHIZOSACCHAROMYCES-POMBE [J].
ATKINS, D ;
IZANT, JG .
CURRENT GENETICS, 1995, 28 (06) :585-588
[3]  
Ausubel F.M., 1996, CURRENT PROTOCOLS MO
[4]  
Bi EF, 1996, MOL CELL BIOL, V16, P5264
[5]   LARGE-SCALE ANALYSIS OF GENE-EXPRESSION, PROTEIN LOCALIZATION, AND GENE DISRUPTION SACCHAROMYCES-CEREVISIAE [J].
BURNS, N ;
GRIMWADE, B ;
ROSSMACDONALD, PB ;
CHOI, EY ;
FINBERG, K ;
ROEDER, GS ;
SNYDER, M .
GENES & DEVELOPMENT, 1994, 8 (09) :1087-1105
[6]   INFLUENCE OF THE CENTROSOME ON THE STRUCTURE OF NUCLEATED MICROTUBULES [J].
EVANS, L ;
MITCHISON, T ;
KIRSCHNER, M .
JOURNAL OF CELL BIOLOGY, 1985, 100 (04) :1185-1191
[7]   COMPARISON OF SCHIZOSACCHAROMYCES-POMBE EXPRESSION SYSTEMS [J].
FORSBURG, SL .
NUCLEIC ACIDS RESEARCH, 1993, 21 (12) :2955-2956
[8]   CELL CYCLE-DEPENDENT SPECIFIC POSITIONING AND CLUSTERING OF CENTROMERES AND TELOMERES IN FISSION YEAST [J].
FUNABIKI, H ;
HAGAN, I ;
UZAWA, S ;
YANAGIDA, M .
JOURNAL OF CELL BIOLOGY, 1993, 121 (05) :961-976
[9]   THE PRODUCT OF THE SPINDLE FORMATION GENE SAD1(+) ASSOCIATES WITH THE FISSION YEAST SPINDLE POLE BODY AND IS ESSENTIAL FOR VIABILITY [J].
HAGAN, I ;
YANAGIDA, M .
JOURNAL OF CELL BIOLOGY, 1995, 129 (04) :1033-1047
[10]   GENETICS OF THE FISSION YEAST SCHIZOSACCHAROMYCES-POMBE [J].
HAYLES, J ;
NURSE, P .
ANNUAL REVIEW OF GENETICS, 1992, 26 :373-402