Germ line transformation of the yellow fever mosquito, Aedes aegypti, mediated by transpositional insertion of a piggyBac vector

被引:47
作者
Lobo, NF [1 ]
Hua-Van, A [1 ]
Li, X [1 ]
Nolen, BM [1 ]
Fraser, MJ [1 ]
机构
[1] Univ Notre Dame, Dept Biol Sci, Ctr Trop Dis Res & Training, Notre Dame, IN 46556 USA
关键词
D O I
10.1046/j.1365-2583.2002.00317.x
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
Mosquito-vectored diseases such as yellow fever and dengue fever continue to have a substantial impact on human populations world-wide. Novel strategies for control of these mosquito vectored diseases can arise through the development of reliable systems for genetic manipulation of the insect vector. A piggyBac vector marked with the Drosophila melanogaster cinnabar (cn) gene was used to transform the white-eyed kh(w) strain of Aedes aegypti . Microinjection of preblastoderm embryos resulted in four families of cinnabar transformed insects. An overall transformation frequency of 4%, with a range of 0% to as high as 13% for individual experiments, was achieved when using a heat-shock induced transposase providing helper plasmid. Southern hybridizations indicated multiple insertion events in three of four transgenic lines, while the presence of duplicated target TTAA sites at either ends of individual insertions confirmed characteristic piggyBac transposition events in these three transgenic lines. The transgenic phenotype has remained stable for more than twenty generations. The transformations effected using the piggyBac element establish the potential of this element as a germ-line transformation vector for Aedine mosquitoes.
引用
收藏
页码:133 / 139
页数:7
相关论文
共 45 条
[1]  
BHALLA SC, 1968, GENETICS, V58, P249
[2]   TRANSPOSON MUTAGENESIS OF BACULOVIRUSES - ANALYSIS OF TRICHOPLUSIA-NI TRANSPOSON IFP2 INSERTIONS WITHIN THE FP-LOCUS OF NUCLEAR POLYHEDROSIS VIRUSES [J].
CARY, LC ;
GOEBEL, M ;
CORSARO, BG ;
WANG, HG ;
ROSEN, E ;
FRASER, MJ .
VIROLOGY, 1989, 172 (01) :156-169
[3]   Stable germline transformation of the malaria mosquito Anopheles stephensi [J].
Catteruccia, F ;
Nolan, T ;
Loukeris, TG ;
Blass, C ;
Savakis, C ;
Kafatos, FC ;
Crisanti, A .
NATURE, 2000, 405 (6789) :959-962
[4]   Mariner transposition and transformation of the yellow fever mosquito, Aedes aegypti [J].
Coates, CJ ;
Jasinskiene, N ;
Miyashiro, L ;
James, AA .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (07) :3748-3751
[5]   Promoter-directed expression of recombinant fire-fly luciferase in the salivary glands of Hermes-transformed Aedes aegypti [J].
Coates, CJ ;
Jasinskiene, N ;
Pott, GB ;
James, AA .
GENE, 1999, 226 (02) :317-325
[6]   Transient expression of the Drosophila melanogaster cinnabar gene rescues eye color in the white eye (WE) strain of Aedes aegypti [J].
Cornel, AJ ;
Benedict, MQ ;
Rafferty, CS ;
Howells, AJ ;
Collins, FH .
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY, 1997, 27 (12) :993-997
[7]   PCR analysis of insertion site specificity, transcription, and structural uniformity of the Lepidopteran transposable element IFP2 in the TN-368 cell genome [J].
Elick, TA ;
Bauser, CA ;
Principe, NM ;
Fraser, MJ .
GENETICA, 1996, 97 (02) :127-139
[8]   Excision of the piggyBac transposable element in vitro is a precise event that is enhanced by the expression of its encoded transposase [J].
Elick, TA ;
Bauser, CA ;
Fraser, MJ .
GENETICA, 1996, 98 (01) :33-41
[9]   HIGH-FREQUENCY P-ELEMENT LOSS IN DROSOPHILA IS HOMOLOG DEPENDENT [J].
ENGELS, WR ;
JOHNSONSCHLITZ, DM ;
EGGLESTON, WB ;
SVED, J .
CELL, 1990, 62 (03) :515-525
[10]   MINOS, A NEW TRANSPOSABLE ELEMENT FROM DROSOPHILA-HYDEI, IS A MEMBER OF THE TC1-LIKE FAMILY OF TRANSPOSONS [J].
FRANZ, G ;
SAVAKIS, C .
NUCLEIC ACIDS RESEARCH, 1991, 19 (23) :6646-6646