Isolation and Transient Expression Assay of PEAMT Gene Promoter from Salicornia europaea
- VernacularTitle:盐角草磷酸乙醇胺N-甲基转移酶基因(SePEAMT)启动子的克隆及瞬时表达
- Author:
Sai-Jian MA
;
Qiao SU
;
Song WU
;
Li-Jia AN
;
- Publication Type:Journal Article
- Keywords:
Phosphoethanolamine N-methyltransferase Anchored PCR Promoter sequence analysis Transient expression
- From:
China Biotechnology
2006;0(08):-
- CountryChina
- Language:Chinese
-
Abstract:
Phosphoethanolamine N-methyltransferase (PEAMT) is a key enzyme that catalyzes the synthesis of phosphocholine, which is an important precursor of phosphatidylcholine and glycine betaine. A 1249bp 5'-flanking region of phosphoethanolamine N-methyltransferase gene was isolated by anchored PCR, based on the cDNA sequence of PEAMT from halophyte Salicornia europaea. The transcription start site was identified as A and localized at 301bp upstream of the ATG according to the results of RLM-RACE. In SePEAMT promoter region, many potential cis-acting elements were predicted by PlantCARE and PLACE programs. Aside from the basal transcriptional elements TATA-box and CAAT-box, some stress-responsive motifs such as ABRE, HSE and LTR were found. In addition, some pollen-specific activation-related elements were also present in this region. Binary expression vector was constructed by fusing SePEAMT promoter with GUS gene and designated as pPro. The pPro was transferred into tobacco by Agrobacterium-medicated transformation and transient GUS expression analysis indicated that SePEAMT promoter could drive strong GUS expression.
