Isolation of a maize ZmCI-1B promoter and characterization of its activity in transgenic maize and tobacco.
The 2-kb ZmCI - 1B promoter is active in the root and embryo and induced by wounding in maize and the 220-bp 5'-deleted segment maybe the minimal promoter. The subtilisin-chymotrypsin inhibitor gene, CI-1B of Zea mays (ZmCI-1B), has been suggested to induce the maize defense system to resist insect attack. Real-time RT-PCR showed that ZmCI-1B gene exhibited especially high expression in roots and embryos. The 2-kb full-length promoter of ZmCI-1B gene was isolated from the maize genome and used to drive expression of a beta-glucuronidase (GUS) reporter gene for transient expression and stable expression analysis in maize. The results of GUS histochemical staining in transgenic maize plants revealed that the ZmCI-1B promoter induced GUS expression preferentially in roots and embryos and in response to wounding. A series of 5'-deleted segments of the ZmCI-1B promoter were cloned individually to drive GUS expression for further analysis. Deletion analysis combined with the histochemical staining of transgenic tobacco plants revealed 220-bp segment could drive GUS in a tissue-specific and wounding-induced expression in tobacco; thus, it maybe the minimally active promoter of ZmCI-1B gene. Furthermore, it revealed that the ZmCI-1B promoter contained tissue-specific and wounding-induced elements.
Department of Crop Genomics and Genetic Improvement, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, 12 ZhongGuanCun South Street, Beijing, 100081, China.
Ye
Y
Li
Xiaoqing
X
Liu
Jie
J
Li
Suzhen
S
Li
Guanyu
G
Chen
Xiaojin
X
Zhou
Wenzhu
W
Yang
Rumei
R
Chen
D002918:Chymotrypsin / Q000037:antagonists & inhibitors / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics / Q000502:physiology; D005966:Glucuronidase / Q000096:biosynthesis / Q000502:physiology; D010940:Plant Proteins / Q000235:genetics* / Q000502:physiology; D018517:Plant Roots / Q000502:physiology; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics* / Q000502:physiology; D060888:Real-Time Polymerase Chain Reaction; D020860:Subtilisin / Q000037:antagonists & inhibitors / Q000235:genetics; D014026:Nicotiana / Q000235:genetics* / Q000502:physiology; D003313:Zea mays / Q000235:genetics* / Q000502:physiology
D010940:Plant Proteins; D005966:Glucuronidase; D002918:Chymotrypsin; D020860:Subtilisin