摘要
将PCR扩增获得的水稻花药绒毡层特异表达基因Osg6B启动子(简写成6B)与来自质粒pROKⅡ上的NOS终止子相连,产生了pGEM7Z-6BNOS。然后将来自解淀粉芽孢杆菌(Bacillus amylodique faciens)中的Barnase和Barstar基因(简写成BN和BS)分别插入到pGEM7Z-6BNO上的BamH I和Nco I位点上,再用Sal I和Xho I切下2.3 kb的6BBNNOS相2.2kb的6BBSNOS片段,并分别将其插入到pDM302的Xho I位点上,经酶切分析表明6BBNNOS和6BBSNOS在pDM302上的方向相反。从而成功地构建了水稻雄性不育和其育性恢复表达载体pDM302-6BBNNOS和pDM302-6BBSNOS。用基因枪轰击水稻幼胚,获得了抗膦化麦黄酮(PPT)的水稻转基因植株,可望形成优良的水稻雄性不育和育性恢复系,以应用于生产。
The 1.7kb promoter 6B of tapetum-specific genes from Oryza sativa L. by
PCR was connected to the 270 bp NOS terminator from pROK II to generate pGEM7Z-6BNOS.
The genes for Barnase and Barstar from Bacillus amylolique faciens were inserted between the
BamH I and Nco I sites of pGEM7Z-6BNOS to give pGEM7Z-6BBNNOS and pGEM7Z-6BB-
NOS, respectively. The fragments of 2.3kb 6BBNNOS and 2. 2kb 6BBSNOS were separated
from pGEM7Z-6BBNNOS and pGEM7Z-6BBSNOS with the Sal I and Xho I digestion, and
cloned into the Xho I sites of pDM302 conferred resistance to phosphinothricin(PPT) , respec
tively. The orientation of the 6BBNNOS and 6BBSNOS fragments was diverse in pDM302.
Therefore, the plant expression vectors of male sterility and its fertility restoration were success
fully constructed. The immatured embryos from rice were transformed by particle bombard
ment. The rice transgcnic plants with resistance to PPT were generated. The superior male ster
ile line and its restorer are being expected.
出处
《作物学报》
CAS
CSCD
北大核心
1998年第5期629-634,共6页
Acta Agronomica Sinica
