RNA DELIVERY IN SACCHAROMYCES-CEREVISIAE USING ELECTROPORATION

被引：24

作者：

EVERETT, JG ^{[1
]}

GALLIE, DR ^{[1
]}

机构：

[1] UNIV CALIF RIVERSIDE, DEPT BIOCHEM, RIVERSIDE, CA 92521 USA

来源：

YEAST | 1992年 / 8卷 / 12期

关键词：

ELECTROPORATION; SPHEROPLASTS; MESSENGER RNA; TRANSLATION;

D O I：

10.1002/yea.320081203

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：

An efficient delivery method for introducing in vitro synthesized RNA into yeast has been developed using electroporation. Spheroplast preparation, electroporation, and subsequent expression analysis can be accomplished within a single day. The use of introduced mRNA constructs avoids any complications due to nuclear regulation and is particularly suited for cytoplasmic regulatory studies. Moreover, this technique is useful for introducing those RNAs that cannot be made in vivo, such as poly(A)- mRNAs or RNAs with base modifications. We demonstrate that the Escherichia coli GUS gene and the firefly Luc gene are both excellent reporter genes for RNA electroporation.

引用

页码：1007 / 1014

页数：8

共 34 条

[1] THE 5'-LEADER SEQUENCE OF TOBACCO MOSAIC-VIRUS RNA MEDIATES INITIATION-FACTOR-4E-INDEPENDENT, BUT STILL INITIATION-FACTOR-4A-DEPENDENT TRANSLATION IN YEAST EXTRACTS [J].

ALTMANN, M ;

BLUM, S ;

WILSON, TMA ;

TRACHSEL, H .

GENE, 1990, 91 (01) :127-129

[2] TRANSLATION IN SACCHAROMYCES-CEREVISIAE - INITIATION-FACTOR 4E-DEPENDENT CELL-FREE SYSTEM [J].

ALTMANN, M ;

SONENBERG, N ;

TRACHSEL, H .

MOLECULAR AND CELLULAR BIOLOGY, 1989, 9 (10) :4467-4472

[3] MESSENGER-RNA CAP-BINDING PROTEIN - CLONING OF THE GENE ENCODING PROTEIN-SYNTHESIS INITIATION FACTOR-EIF-4E FROM SACCHAROMYCES-CEREVISIAE [J].

ALTMANN, M ;

HANDSCHIN, C ;

TRACHSEL, H .

MOLECULAR AND CELLULAR BIOLOGY, 1987, 7 (03) :998-1003

[4] THE NUMBER OF MOLECULES TAKEN UP BY ELECTROPORATED CELLS - QUANTITATIVE-DETERMINATION [J].

BARTOLETTI, DC ;

HARRISON, GI ;

WEAVER, JC .

FEBS LETTERS, 1989, 256 (1-2) :4-10

[5]

BECKER DM, 1991, METHOD ENZYMOL, V194, P182

[6] TRANSLATION IN SACCHAROMYCES-CEREVISIAE - INITIATION-FACTOR 4A-DEPENDENT CELL-FREE SYSTEM [J].

BLUM, S ;

MUELLER, M ;

SCHMID, SR ;

LINDER, P ;

TRACHSEL, H .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (16) :6043-6046

[7]

CALLIS J, 1987, NUCLEIC ACIDS RES, V15, P5823

[8] BETA-GLOBIN MESSENGER-RNAS CAPPED WITH M7G, M2(2),7G OR M3(2)2,7G DIFFER IN INTRINSIC TRANSLATION EFFICIENCY [J].

DARZYNKIEWICZ, E ;

STEPINSKI, J ;

EKIEL, I ;

JIN, Y ;

HABER, D ;

SIJUWADE, T ;

TAHARA, SM .

NUCLEIC ACIDS RESEARCH, 1988, 16 (18) :8953-8962

[9] FIREFLY LUCIFERASE GENE - STRUCTURE AND EXPRESSION IN MAMMALIAN-CELLS [J].

DEWET, JR ;

WOOD, KV ;

DELUCA, M ;

HELINSKI, DR ;

SUBRAMANI, S .

MOLECULAR AND CELLULAR BIOLOGY, 1987, 7 (02) :725-737

[10]

FROMM M, 1987, METHOD ENZYMOL, V153, P351

← 1 2 3 4 →