One-step PCR mediated strategy for the construction of conditionally expressed and epitope tagged yeast proteins

被引：84

作者：

Lafontaine, D ^{[1
]}

Tollervey, D ^{[1
]}

机构：

[1] EUROPEAN MOL BIOL LAB,D-69012 HEIDELBERG,GERMANY

来源：

NUCLEIC ACIDS RESEARCH | 1996年 / 24卷 / 17期

关键词：

D O I：

10.1093/nar/24.17.3469

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

With the availability of the complete yeast genomic sequence, techniques which allow the rapid functional analysis of genes of interest are of increasing importance. Here we report a technique which allows the initial characterisation of genes of interest, through the construction of conditionally expressed mutations for functional analyses and the generation of epitope-tagged fusion proteins for immuno-localisation and immuno-purification, entirely by PCR.

引用

页码：3469 / 3471

页数：3

共 9 条

[1] A SIMPLE AND EFFICIENT METHOD FOR DIRECT GENE DELETION IN SACCHAROMYCES-CEREVISIAE
BAUDIN, A
OZIERKALOGEROPOULOS, O
DENOUEL, A
LACROUTE, F
CULLIN, C
[J]. NUCLEIC ACIDS RESEARCH, 1993, 21 (14) : 3329 - 3330
[2] CLARKE MW, 1990, J CELL BIOL, V11, P1741
[3] IMPROVED METHOD FOR HIGH-EFFICIENCY TRANSFORMATION OF INTACT YEAST-CELLS
GIETZ, D
STJEAN, A
WOODS, RA
SCHIESTL, RH
[J]. NUCLEIC ACIDS RESEARCH, 1992, 20 (06) : 1425 - 1425
[4] THE 18S RIBOSOMAL-RNA DIMETHYLASE DIM1P IS REQUIRED FOR PRE-RIBOSOMAL-RNA PROCESSING IN YEAST
LAFONTAINE, D
VANDENHAUTE, J
TOLLERVEY, D
[J]. GENES & DEVELOPMENT, 1995, 9 (20) : 2470 - 2481
[5] A METHOD FOR PERFORMING PRECISE ALTERATIONS IN THE YEAST GENOME USING A RECYCLABLE SELECTABLE MARKER
LANGLEROUAULT, F
JACOBS, E
[J]. NUCLEIC ACIDS RESEARCH, 1995, 23 (15) : 3079 - 3081
[6] LARGE-SCALE AFFINITY PURIFICATION OF HUMAN INSULIN-LIKE GROWTH FACTOR-I FROM CULTURE-MEDIUM OF ESCHERICHIA-COLI
MOKS, T
ABRAHMSEN, L
OSTERLOF, B
JOSEPHSON, S
OSTLING, M
ENFORS, SO
PERSSON, I
NILSSON, B
UHLEN, M
[J]. BIO-TECHNOLOGY, 1987, 5 (04): : 379 - 382
[7] A YEAST NUCLEOLAR PROTEIN RELATED TO MAMMALIAN FIBRILLARIN IS ASSOCIATED WITH SMALL NUCLEOLAR RNA AND IS ESSENTIAL FOR VIABILITY
SCHIMMANG, T
TOLLERVEY, D
KERN, H
FRANK, R
HURT, EC
[J]. EMBO JOURNAL, 1989, 8 (13) : 4015 - 4024
[8] SIKORSKI RS, 1989, GENETICS, V122, P19
[9] DELETION ANALYSIS OF THE SACCHAROMYCES GAL GENE-CLUSTER - TRANSCRIPTION FROM 3 PROMOTERS
STJOHN, TP
SCHERER, S
MCDONELL, MW
DAVIS, RW
[J]. JOURNAL OF MOLECULAR BIOLOGY, 1981, 152 (02) : 317 - 334

← 1 →