摘要
利用我们自己分离的甘露碱含成酶基因启动子与萤光素酶结构基因、胭脂碱合成酶基因’3末端结构相拼构成一融合基因,并在带有此融合基因的中间载体PBZ7610插入Ti质粒T区的tmr基因,构成中间载体pBZ7621。利用改建的Ti质粒载体PGV3850,将萤光素酶融合基因引入了烟草植株,结果表明,萤光素酶融合基因在转化烟草中能表达。中间载体pBZ7610还带有PstⅠ,HindⅢ,XbaⅠ等多个单一的酶切位点,外源基因极易插入。利用中间载体pBZ7621,还可研究启动子在高等植物不同发育阶段中的功能特征。
Firefly luciferase gene encodes an enzyme
that catalyzes the light-producing, adenosine
triphosphate (ATP)-dependent oxidation of
luciferin. This gene has been expressed in
bacteria, mammalian and plant cells.we
constructed a chimeric luciferase gene which
was promoted by isolated dual promoter frag-
ment 2' and ended by 3' end of nopaline syn-
thase gene. Using tmr gene which was isolated
from the T-region of Ti plasmid, we set up an
intermediate vector pBZ7621 (Fig. 1). The
chimeric luciferase gene was introduced into
tobacco by using modified Ti plasmid pGV
3850 of Agrobacterium tumefaciens (Fig. 2). The
result shows that the luciferase gene can be
expressed in higher plant (Plate Ⅰ,Ⅱ). The
luciferase gene can be particularly valuable
in plant genetics as a marker for plant cell trans-
formation, as a reporter for studying promoter
function and as a probe for a variety of plant cell
functions.
The intermediate vector pBZ 7610 has not
only the chimeric luciferase gene, but also some
unique restriction enzyme sites like Xba I, Pst
Ⅰ, Hind Ⅲ and so on, into which the foreign
genes can be easily inserted. Using pBZ 7621,
one can study the function of any promoter
during the development of higher plants.
关键词
启动子
萤光素酶基因
烟草
转化
luciferase gene
tobacco
transformation
expression
