Establishment of an efficient BAC transgenesis protocol and its application to functional characterization of the mouse Brachyury locus

被引:24
作者
Abe, K
Hazama, M
Katoh, H
Yamamura, K
Suzuki, M
机构
[1] RIKEN, Tsukuba Inst, BioResource Ctr, Res & Dev Team Mammalian Cellular Dynam, Tsukuba, Ibaraki 3050074, Japan
[2] Kyoto Univ, Fac Med, Dept Biol Sci, Sakyo Ku, Kyoto 6068501, Japan
[3] Hamamatsu Univ Sch Med, Expt Anim Inst, Shizuoka 4313192, Japan
[4] Kumamoto Univ, Inst Mol Embryol & Genet, Kumamoto 8620976, Japan
[5] Kumamoto Univ, Ctr Anim Res & Dev, Kumamoto 8600811, Japan
关键词
BAC; development; T-locus; transgenesis;
D O I
10.1538/expanim.53.311
中图分类号
S85 [动物医学(兽医学)];
学科分类号
0906 ;
摘要
Transgenesis using large DNA such as YAC or BAC has extended the range of applications in functional genomics. Here we describe an efficient BAC transgenesis protocol using a simple BAC DNA preparation method adopted from YAC DNA purification methods. This method allowed us to isolate BAC DNA from small scale culture of BAC-containing cells in sufficient quantity and purity for microinjection. More than 40 founders have been produced with linearized BAC DNA prepared by this method, and 85% of them contained intact BAC transgenes. In contrast, when circular BAC DNA was injected, an approximately three-fold reduction of transgene integration rate was observed and fewer intact transgene integrations were obtained. A line of transgenic mice carrying a 170-kb BAC clone generated in this way successfully rescued tail and embryonic lethality phenotypes of the mouse Brachyury (T) mutants, further demonstrating the utility of this method in functional analysis of the mouse genome.
引用
收藏
页码:311 / 320
页数:10
相关论文
共 24 条
[1]   Molecular and embryological characterization of a new transgene-induced null allele of mouse Brachyury locus [J].
Abe, K ;
Yamamura, K ;
Suzuki, P .
MAMMALIAN GENOME, 2000, 11 (03) :238-240
[2]   Functional identification of the mouse circadian Clock gene by transgenic BAC rescue [J].
Antoch, MP ;
Song, EJ ;
Chang, AM ;
Vitaterna, MH ;
Zhao, YL ;
Wilsbacher, LD ;
Sangoram, AM ;
King, DP ;
Pinto, LH ;
Takahashi, JS .
CELL, 1997, 89 (04) :655-667
[3]  
BENNETT D, 1975, CELL, V28, P463
[4]  
BENNETT DOROTHEA, 1958, J MORPH, V103, P135, DOI 10.1002/jmor.1051030106
[5]   Development of the short-tailed mutant in the house mouse [J].
Chesley, P .
JOURNAL OF EXPERIMENTAL ZOOLOGY, 1935, 70 (03) :429-459
[6]   Linearization and purification of BAC DNA for the development of transgenic mice [J].
Chrast, R ;
Scott, HS ;
Antonarakis, SE .
TRANSGENIC RESEARCH, 1999, 8 (02) :147-150
[7]   Genome-wide, large-scale production of mutant mice by ENU mutagenesis [J].
de Angelis, MH ;
Flaswinkel, H ;
Fuchs, H ;
Rathkolb, B ;
Soewarto, D ;
Marschall, S ;
Heffner, S ;
Pargent, W ;
Wuensch, K ;
Jung, M ;
Reis, A ;
Richter, T ;
Alessandrini, F ;
Jakob, T ;
Fuchs, E ;
Kolb, H ;
Kremmer, E ;
Schaeble, K ;
Rollinski, B ;
Roscher, A ;
Peters, C ;
Meitinger, T ;
Strom, T ;
Steckler, T ;
Holsboer, F ;
Klopstock, T ;
Gekeler, F ;
Schindewolf, C ;
Jung, T ;
Avraham, K ;
Behrendt, H ;
Ring, J ;
Zimmer, A ;
Schughart, K ;
Pfeffer, K ;
Wolf, E ;
Balling, R .
NATURE GENETICS, 2000, 25 (04) :444-447
[8]   Size matters: Use of YACs, BACs and PACs in transgenic animals [J].
Giraldo, P ;
Montoliu, L .
TRANSGENIC RESEARCH, 2001, 10 (02) :83-103
[9]   Different regulatory sequences are required for parvalbumin gene expression in skeletal muscles and neuronal cells of transgenic mice [J].
Hazama, M ;
Watanabe, D ;
Suzuki, M ;
Mizoguchi, A ;
Pastan, I ;
Nakanishi, S .
MOLECULAR BRAIN RESEARCH, 2002, 100 (1-2) :53-66
[10]  
Herrmann Bernhard G., 1995, Seminars in Developmental Biology, V6, P385, DOI 10.1016/S1044-5781(06)80002-2