Functional analysis of BADH gene promoter from Suaeda liaotungensis K.
A 1,993 bp region upstream of the gene encoding the betaine aldehyde dehydrogenase (BADH) was isolated from Suaeda liaotungensis K., and the analysis of the promoter sequence has revealed the existence of several putative cis-elements by the PLACE database. In this study, according to the characteristic of the BADH promoter, five chimeric constructs varied in the length of promoter fragments from -1,993, -1,466, -1,084, -573 and -300 to +62 bp relative to the transcriptional start site were placed to the upstream of the beta-glucuronidase (GUS) coding region and transferred to Nicotiana tabacum L.cv.89 by Agrobacterium tumefaciens-mediated leaf-disc transformation. The functional properties of each promoter fragment were examined by GUS histochemical staining and fluorescence quantitative analyses in the transgenic tobacco leaves treated with different NaCl concentrations for 48 h. The results show that healthy transgenic plants had decreased GUS activity in leaves, whereas a higher GUS activity was observed when the transgenic plants were challenged with sodium chloride (NaCl). Induction levels were proportional to the concentration of NaCl treatment, allowing fine-tuning of protein expression. GUS enzyme activity was enhanced 6.3-fold in transgenic tobacco leaves containing -300 bp promoter fragment in the presence of 400 mmol/l NaCl compared to the noninductive leaves. This suggests that the smallest promoter fragment (-300 to +62 bp) possesses all the essential cis-acting elements and is sufficient for NaCl induction.
Zhang
Yi
Y
College of Life Sciences, Liaoning Normal University, Dalian, Liaoning, China. zhangyi_81@126.com
Yin
Hui
H
Li
Dan
D
Zhu
Weiwei
W
Li
Qiuli
Q
D001483:Base Sequence; D050728:Betaine-Aldehyde Dehydrogenase / Q000235:genetics*; D019663:Chenopodiaceae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000235:genetics; D030821:Plants, Genetically Modified; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012965:Sodium Chloride / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics
D010940:Plant Proteins; D012965:Sodium Chloride; D050728:Betaine-Aldehyde Dehydrogenase; D005966:Glucuronidase