BARLEY ALEURONE LAYER CELL PROTOPLASTS AS A TRANSIENT EXPRESSION SYSTEM

被引：15

作者：

GOPALAKRISHNAN, B ^{[1
]}

SONTHAYANON, B ^{[1
]}

RAHMATULLAH, R ^{[1
]}

MUTHUKRISHNAN, S ^{[1
]}

机构：

[1] KANSAS STATE UNIV AGR & APPL SCI,DEPT BIOCHEM,WILLARD HALL,MANHATTAN,KS 66506

来源：

PLANT MOLECULAR BIOLOGY | 1991年 / 16卷 / 03期

关键词：

ALEURONE PROTOPLASTS; BARLEY; HORMONE-RESPONSIVE ALPHA-AMYLASE SYNTHESIS; TRANSIENT EXPRESSION;

D O I：

10.1007/BF00023996

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Protoplasts were prepared from barley aleurone layers using 'Onozuka' cellulase digestion and purification through a Percoll gradient. Protoplasts prepared by this procedure had a viability ranging from 60% to 80% during the first two days of culture. They were responsive to gibberellic acid (GA) as measured by the stimulation of alpha-amylase synthesis. The GA stimulation was counteracted by abscisic acid (ABA). In the presence of polyethylene glycol (PEG), the protoplasts took up exogenously added plasmid DNA containing the reporter gene coding for chloramphenicol acetyl transferase (CAT) linked to a 35S promoter from cauliflower mosaic virus (CaMV) or to barley alpha-amylase gene promoters and expressed CAT activity. Therefore, barley aleurone layer protoplasts are suitable for analysis of hormone-responsive elements in hydrolase genes.

引用

页码：463 / 467

页数：5

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