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Cloning and Plant Expression Vector Construction of AtLACS9 Gene from Arabidopsis Thaliana(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

Issue:
2011年02期
Page:
190-193
Research Field:
Publishing date:

Info

Title:
Cloning and Plant Expression Vector Construction of AtLACS9 Gene from Arabidopsis Thaliana
Author(s):
ZHAO Huan-huan;  WU Xing;  ZHANG Feng;  LI Hong-wei; WANG Mao-yan
College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018，Inner Mongolia, China
Keywords:
Long-chain acyl-CoA synthetases (LACSs); Gene cloning; Vector construction
PACS:
S565.1
DOI:
10.11861/j.issn.1000-9841.2011.02.0190
Abstract:
Triacylglycerol (TAG) is the major form of storage lipid in oilseeds, and acyl-coenzyme A (CoA) and glycerol-3-phosphate are precursors for the synthesis of TAG. The long-chain acyl-CoA synthetases (LACSs) catalyze the synthesis of acyl-CoA molecules and therefore influence TAG content in oilseeds. In this paper, the soybean seed-specific lectin promoter(lec), which was digested with restriction enzymes from vector pBI-lec, was inserted into the multiple cloning site of the plant expression vector pCAMBIA 3301(p3301), and then the complete coding region cDNA of Arabidopsis thaliana LACS9(AtLACS9) gene was amplified by reverse transcription PCR（RT-PCR）and subcloned into the reconstructed vector p3301 downstream of the lec promoter. This work aimed at laying a foundation for future transformation of soybean with the gene and then obtaining transgenic soybean with enhanced oil content. The results showed that the vector p3301 was successully reconstructed by inserting the lec promoter and was named p3301-lec; AtLACS9 had an open reading frame of 2076 bp and the corresponding protein consists of 691 amino acids; the seed-specific expression vector of AtLACS9, namely p3301-lec-AtLACS9, was constructed successfully and could be used in the next experiments.

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Last Update: 2014-09-11