Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.15 (pectinase)
 2,440 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

This report describes a method for the easy generation of inverted repeat constructs for the silencing of genes of unknown sequence which is applicable to high-throughput studies. This improved procedure for high-efficiency gene silencing is specific for a target gene, but does not require inverted repeat DNA of the target gene in the construct. The method employs an inverted repeat of the 3'-untranslated region (3'-UTR) of a heterologous gene, and has been demonstrated using the 3'-UTR region of the nopaline synthase (nos) gene from Agrobacterium tumefaciens, which is often used as the 3'-UTR for transgene constructs. In a population of independent tomato primary transformants harboring a stably integrated polygalacturonase (PG) transgene driven by a constitutive promoter and linked to an inverted repeat of the nos 3'-UTR, 51 of 56 primary transformants (91% of the population) showed highly effective post-transcriptional silencing of the PG gene, with PG mRNA abundance in ripe fruit reduced by 98% or more. The method was also effective in Arabidopsis, where two different, relatively uncharacterized plant transcription factors were also targeted effectively. This method has the advantage of ease and rapidity in preparation of the constructs, since a gene of interest can be inserted into a binary vector already containing the promoter and the inverted nos domain in a single-cloning step, and does not require any knowledge of the DNA sequence. The approach is suitable for high-throughput gene silencing studies, where it is necessary to investigate the function of hundreds to thousands of uncharacterized genes.
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PMID:Inverted repeat of a heterologous 3'-untranslated region for high-efficiency, high-throughput gene silencing. 1260 50

As with many phytopathogenic bacteria, the virulence of Xanthomonas campestris pv. campestris (Xcc), the causal agent of black rot disease in cruciferous plants, relies on secretion of a suite of extracellular enzymes that includes cellulase (endoglucanase), pectinase, protease and amylase. Although the role in virulence of a number of these enzymes has been assessed, the contribution of amylase to Xcc virulence has yet to be established. In this work, we investigated both the role of extracellular amylase in Xcc virulence and the control of its expression. Deletion of XC3487 (here renamed amyAXcc), a putative amylase-encoding gene from the genome of Xcc strain 8004, resulted in a complete loss of extracellular amylase activity and significant reduction in virulence. The extracellular amylase activity and virulence of the amyAXcc mutant could be restored to the wild-type level by expressing amyAXcc in trans. These results demonstrated that amyAXcc is responsible for the extracellular amylase activity of Xcc, and indicated that extracellular amylase plays an important role in Xcc virulence. We further found that the expression of amyAXcc is strongly induced by starch and requires activation by the global post-transcriptional regulator RsmA. RsmA binds specifically to the 5'-untranslated region (5'UTR) of amyAXcc transcripts, suggesting that RsmA regulates amyAXcc directly at the post-transcriptional level. Unexpectedly, in addition to post-transcriptional regulation, the use of a transcriptional reporter demonstrated that RsmA also regulates amyAXcc expression at the transcriptional level, possibly by an indirect mechanism.
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PMID:Extracellular amylase is required for full virulence and regulated by the global post-transcriptional regulator RsmA in Xanthomonas campestris pathovar campestris. 3324 53