EFFICIENT GENERATION OF INFECTIOUS RECOMBINANT BACULOVIRUSES BY SITE-SPECIFIC TRANSPOSON-MEDIATED INSERTION OF FOREIGN GENES INTO A BACULOVIRUS GENOME PROPAGATED IN ESCHERICHIA-COLI

被引:704
作者
LUCKOW, VA [1 ]
LEE, SC [1 ]
BARRY, GF [1 ]
OLINS, PO [1 ]
机构
[1] MONSANTO CO, AGR GRP, CHESTERFIELD, MO 63198 USA
关键词
D O I
10.1128/JVI.67.8.4566-4579.1993
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The construction and purification of recombinant baculovirus vectors for the expression of foreign genes in insect cells by standard transfection and plaque assay methods can take as long as 4 to 6 weeks. This period can be reduced to several days by using a novel baculovirus shuttle vector (bacmid) that can replicate in Escherichia coli as a plasmid and can infect susceptible lepidopteran insect cells. The bacmid is a recombinant virus that contains a mini-F replicon, a kanamycin resistance marker, and attTn7, the target site for the bacterial transposon Tn7. Expression cassettes comprising a baculovirus promoter driving expression of a foreign gene that is flanked by the left and right ends of Tn7 can transpose to the target bacmid in E. coli when Tn7 transposition functions are provided in trans by a helper plasmid. The foreign gene is expressed when the resulting composite bacmid is introduced into insect cells.
引用
收藏
页码:4566 / 4579
页数:14
相关论文
共 58 条
[1]   PBLUESCRIPT-II - GENE-MAPPING VECTORS [J].
ALTINGMEES, MA ;
SHORT, JM .
NUCLEIC ACIDS RESEARCH, 1989, 17 (22) :9494-9494
[3]   PERMANENT INSERTION OF FOREIGN GENES INTO THE CHROMOSOMES OF SOIL BACTERIA [J].
BARRY, GF .
BIO-TECHNOLOGY, 1986, 4 (05) :446-449
[4]  
BECKER DM, 1991, METHOD ENZYMOL, V194, P182
[5]  
BERG CM, 1989, MOBILE DNA, P879
[6]   SUPERPOLYLINKERS IN CLONING AND EXPRESSION VECTORS [J].
BROSIUS, J .
DNA-A JOURNAL OF MOLECULAR & CELLULAR BIOLOGY, 1989, 8 (10) :759-777
[7]   SCREENING RECOMBINANT BACULOVIRUS PLAQUES INSITU WITH ANTIBODY PROBES [J].
CAPONE, J .
GENE ANALYSIS TECHNIQUES, 1989, 6 (03) :62-66
[8]   ONE-STEP PREPARATION OF COMPETENT ESCHERICHIA-COLI - TRANSFORMATION AND STORAGE OF BACTERIAL-CELLS IN THE SAME SOLUTION [J].
CHUNG, CT ;
NIEMELA, SL ;
MILLER, RH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1989, 86 (07) :2172-2175
[9]   RECOGNITION AND CLEAVAGE SITE OF THE INTRON-ENCODED OMEGA TRANSPOSASE [J].
COLLEAUX, L ;
DAURIOL, L ;
GALIBERT, F ;
DUJON, B .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (16) :6022-6026
[10]  
CRAIG NL, 1989, MOBILE DNA, P211