Expression enhancing intron sequences

摘要

The invention relates to methods for the identification and use of introns with gene expression enhancing properties. The teaching of this invention enables the identification of introns causing intron-mediated enhancement (IME) of gene expression. The invention furthermore relates to recombinant expression construct and vectors comprising said IME-introns operably linked with a promoter sequence and a nucleic acid sequence. The present invention also relates to transgenic plants and plant cells transformed with these recombinant expression constructs or vectors, to cultures, parts or propagation material derived there from, and to the use of same for the preparation of foodstuffs, animal feeds, seed, pharmaceuticals or fine chemicals, to improve plant biomass, yield, or provide desirable phenotypes.

主权项

1. A recombinant DNA expression construct comprising:
a) at least one promoter sequence functioning in plants or plant cells; b) at least one intron comprising the sequence of SEQ ID NO: 2 or a functional equivalent thereof; and c) at least one nucleic acid sequence,wherein the at least one promoter sequence and at least one intron are functionally linked to the at least one nucleic acid sequence,wherein the at least one intron is heterologous to the at least one nucleic acid sequence and/or to the at least one promoter sequence, andwherein the functional equivalent thereof increases gene expression and comprises a sequence having at least 80% sequence identity to the full-length sequence of SEQ ID NO: 2 and comprises
I) an intron length shorter than 1000 base pairs, and II) a 5′ splice site comprising the dinucleotide sequence 5′-GT-3′ (SEQ ID NO: 78), and III) a 3′ splice site comprising the trinucleotide sequence 5′-CAG-3′ (SEQ ID NO: 79), and IV) a branch point resembling the consensus sequence 5′-CURAY-3′ (SEQ ID NO: 75) upstream of the 3′ splice site.