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Discovery, cloning and heterologous expression of secreted potato proteins reveal erroneous pre-mRNA splicing in Aspergillus oryzae

Paper ID Volume ID Publish Year Pages File Format Full-Text
25678 43590 2006 12 PDF Available
Title
Discovery, cloning and heterologous expression of secreted potato proteins reveal erroneous pre-mRNA splicing in Aspergillus oryzae
Abstract

A novel transposon assisted signal trapping (TAST) technology, developed to specifically select only the secreted proteins, was used to discover novel extracellular plant proteins from Solarium tuberosum infected with Phytophthora infestans. Analysis of 384 hits provided 191 P. infestans and S. tuberosum sequences of secreted proteins, with an approx. 2/3 of these originating from potato. Subsequent screening for interesting genes was carried out using bioinformatics. A selected variety of the discovered sequences are presented, including a novel S. tuberosum xyloglucan endotransglucosylase (StXTH), which was cloned and subjected to detailed heterologous expression studies in Aspergillus oryzae. RT-PCR analysis of mRNA from A. oryzae StXTH1 transformants revealed that parts of the mRNA pool had been incorrectly processed, and only weak and inconsistent indications of active protein could be detected. A high AT content of StXTH1 and the occurrence of A. oryzae intron donor, acceptor, and branch point recognition sites resulted in erroneous intron interpretation (cryptic introns) of parts of the mRNA coding sequence. This may explain the difficulties generally experienced in expressing plant genes in filamentous fungi.

Keywords
Cryptic introns; Pre-mRNA splicing; Transposon assisted signal trapping (TAST); Secreted proteins; Heterologous expression; Filamentous fungi
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Discovery, cloning and heterologous expression of secreted potato proteins reveal erroneous pre-mRNA splicing in Aspergillus oryzae
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 126, Issue 3, 10 November 2006, Pages 265–276
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us