Isolation and Expression of a Barley β-1, 3-Glucanase Isoenzyme II Gene

Abstract

A β-1, 3-glucanase gene from Hordeum vulgare was isolated by a PCR strategy, cloned and subsequently sequenced. The amplified sequence contained the entire coding region of the isoenzyme II, which is interrupted by a 165 bp intron at 73 bp downstream the starting codon. This intron contains all the elements required for the processing mechanism in monocots : a high A + U content, the appropriate splice sites in the 5′ and 3′ ends and four typical YUNAN consensus sequences. Transient transformation of wheat protoplasts with the complete β-1, 3-glucanase gene under the control of maize polyubiquitin promoter revealed that the intron sequence was spliced out. The gene was also expressed at high levels, probably due to an enhancer-like sequence found near the 3′ end of the intron.

Key Words:

• β-1
• 3-glucanase
• Hordeum vulgare
• monocot intron
• Pathogenesis-related
• transient expression
• wheat protoplasts