Difference between revisions of "Part:BBa K678001"

Revision as of 09:38, 7 September 2011

PalcA, inducible Aspergillus nidulans promoter

The promoter alcA cotrols the expression of the gene alcA encoding alcohol dehydrogenase I (ADHI) in Aspergillus nidulans and is a part of the ethanol utilization regulon. PalcA is a tightly regulated promoter and can completely switch off gene expression (1).

Usage and Biology

This promoter has been documented and described extensively in the literature and has among others previously been employed for confirmation of essentiality of genes and overexpression of homo- and heterologous proteins (1). Expression can be strongly induced by the positive transcriptional regulator AlcR by various substrates with a hydroxide group such as ethanol or threonine (2). Complete repression is achieved in the presence of glucose, where the central catabolite regulator CreA controls transcription (3). Lastly transcriptional derepression occurs in the presence of carbon sources that are poor for A. nidulans such as lactose and glycerol. When induced the promoter is very strong and hence results in a high level of expression under inducing conditions (4).

This promoter has also been shown to drive tightly regulated conditional gene expression in Aspergillus fumigatus. In Aspergillus niger the A. nidulans alcR gene needs to be inserted in order for the A. nidulans alcA gene to be expressed (5).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 274
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K678001 short</partinfo>
-The promoter alcA cotrols the expression of the gene ''alcA'' encoding alcohol dehydrogenase I (ADHI) in ''Aspergillus nidulans'' and is a part of the ethanol utilization regulon. PalcA is a tightly regulated promoter and can completely switch off gene expression.
+The promoter alcA cotrols the expression of the gene ''alcA'' encoding alcohol dehydrogenase I (ADHI) in ''Aspergillus nidulans'' and is a part of the ethanol utilization regulon. PalcA is a tightly regulated promoter and can completely switch off gene expression (1).
@@ Line 8: / Line 8: @@
 ===Usage and Biology===
-This promoter has been documented and described extensively in the literature and has among others previously been employed for confirmation of essentiality of genes and overexpression of homo- and heterologous proteins.
+This promoter has been documented and described extensively in the literature and has among others previously been employed for confirmation of essentiality of genes and overexpression of homo- and heterologous proteins (1).
-Expression can be strongly induced by the positive transcriptional regulator AlcR by various substrates with a hydroxide group such as ethanol or threonine (Creaser et al 1985). Complete repression is achieved in the presence of glucose, where the central catabolite regulator CreA controls transcription (Bailey and Arst, 1975). Lastly transcriptional derepression occurs in the presence of carbon sources that are poor for ''A. nidulans'' such as lactose and glycerol. When induced the promoter is very strong and hence results in a high level of expression under inducing conditions (alc5).
+Expression can be strongly induced by the positive transcriptional regulator AlcR by various substrates with a hydroxide group such as ethanol or threonine (2). Complete repression is achieved in the presence of glucose, where the central catabolite regulator CreA controls transcription (3). Lastly transcriptional derepression occurs in the presence of carbon sources that are poor for ''A. nidulans'' such as lactose and glycerol. When induced the promoter is very strong and hence results in a high level of expression under inducing conditions (4).
-This promoter has also been shown to drive tightly regulated conditional gene expression in ''Aspergillus fumigatus''. In ''Aspergillus niger'' the ''A. nidulans'' ''alcR'' gene needs to be inserted in order for the ''A. nidulans alcA'' gene to be expressed.
+This promoter has also been shown to drive tightly regulated conditional gene expression in ''Aspergillus fumigatus''. In ''Aspergillus niger'' the ''A. nidulans'' ''alcR'' gene needs to be inserted in order for the ''A. nidulans alcA'' gene to be expressed (5).
-We suggest to use threonine as an inducer, because it is not a substrate for ADHI and it results in a strong induction.