Expression plasmid vectors with convenient subcloning sites in lambda gt11 that efficiently produce detectable tagged proteins

被引:5
作者
Takemoto, Y [1 ]
Sato, M [1 ]
Furuta, M [1 ]
Hashimoto, Y [1 ]
机构
[1] SYNTEX ROCHE,INST IMMUNOL,NODA,CHIBA 278,JAPAN
关键词
D O I
10.1089/dna.1997.16.893
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
We have generated cDNA expression vectors that efficiently produce tagged proteins. The newly introduced cloning site of this plasmid facilitates subcloning of cDNA in the lambda gt11 phage into the plasmid vector, Because the cDNA is inserted next to the moths of the tagged DNA sequence, the protein produced by the tag sequence-coupled cDNA is easily detected by Western blot analysis or immunoprecipitation using commercially available antibodies, The double-tagged protein significantly enhances the efficiency of Western blot and immunoprecipitation detection as compared with the single-tagged protein.
引用
收藏
页码:893 / 896
页数:4
相关论文
共 12 条
[1]   MAX - A HELIX-LOOP-HELIX ZIPPER PROTEIN THAT FORMS A SEQUENCE-SPECIFIC DNA-BINDING COMPLEX WITH MYC [J].
BLACKWOOD, EM ;
EISENMAN, RN .
SCIENCE, 1991, 251 (4998) :1211-1217
[2]   ISOLATION OF MONOCLONAL-ANTIBODIES SPECIFIC FOR HUMAN C-MYC PROTO-ONCOGENE PRODUCT [J].
EVAN, GI ;
LEWIS, GK ;
RAMSAY, G ;
BISHOP, JM .
MOLECULAR AND CELLULAR BIOLOGY, 1985, 5 (12) :3610-3616
[3]   PURIFICATION OF A RAS-RESPONSIVE ADENYLYL CYCLASE COMPLEX FROM SACCHAROMYCES-CEREVISIAE BY USE OF AN EPITOPE ADDITION METHOD [J].
FIELD, J ;
NIKAWA, J ;
BROEK, D ;
MACDONALD, B ;
RODGERS, L ;
WILSON, IA ;
LERNER, RA ;
WIGLER, M .
MOLECULAR AND CELLULAR BIOLOGY, 1988, 8 (05) :2159-2165
[4]  
Gorman C., 1985, DNA CLONING PRACTICA, V1, P143
[5]   HUMAN WEE-1 MAINTAINS MITOTIC TIMING BY PROTECTING THE NUCLEUS FROM CYTOPLASMICALLY ACTIVATED CDC2 KINASE [J].
HEALD, R ;
MCLOUGHLIN, M ;
MCKEON, F .
CELL, 1993, 74 (03) :463-474
[6]   QUANTITATIVE-DETERMINATION THAT ONE OF 2 POTENTIAL RNA-BINDING DOMAINS OF THE A-PROTEIN COMPONENT OF THE U1 SMALL NUCLEAR RIBONUCLEOPROTEIN COMPLEX BINDS WITH HIGH-AFFINITY TO STEM LOOP-II OF U1 RNA [J].
LUTZFREYERMUTH, C ;
QUERY, CC ;
KEENE, JD .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (16) :6393-6397
[7]  
MACGREGOR PF, 1990, ONCOGENE, V5, P451
[8]   USE OF PEPTIDE TAGGING TO DETECT PROTEINS EXPRESSED FROM CLONED GENES - DELETION MAPPING FUNCTIONAL DOMAINS OF DROSOPHILA HSP70 [J].
MUNRO, S ;
PELHAM, HRB .
EMBO JOURNAL, 1984, 3 (13) :3087-3093
[9]   CLONING OF PI3 KINASE ASSOCIATED P85 UTILIZING A NOVEL METHOD FOR EXPRESSION CLONING OF TARGET PROTEINS FOR RECEPTOR TYROSINE KINASES [J].
SKOLNIK, EY ;
MARGOLIS, B ;
MOHAMMADI, M ;
LOWENSTEIN, E ;
FISCHER, R ;
DREPPS, A ;
ULLRICH, A ;
SCHLESSINGER, J .
CELL, 1991, 65 (01) :83-90
[10]   LCKBP1, A PROLINE-RICH PROTEIN EXPRESSED IN HEMATOPOIETIC LINEAGE CELLS, DIRECTLY ASSOCIATES WITH THE SH3 DOMAIN OF PROTEIN-TYROSINE KINASE P56(LCK) [J].
TAKEMOTO, Y ;
FURUTA, M ;
LI, XK ;
STRONGSPARKS, WJ ;
HASHIMOTO, Y .
EMBO JOURNAL, 1995, 14 (14) :3403-3414