Assessment of the core cryparin promoter from Cryphonectria parasitica for heterologous expression in filamentous fungi.
Cryparin is an abundant cell-wall-associated hydrophobin of Cryphonectria parasitica. Although cryparin is encoded as a single copy gene, it is the most abundant protein produced by this fungus when grown in liquid culture. Studies to characterize the transcriptional regulatory element(s) found that the fragment between nt -188 and the start codon was the minimal but sufficient promoter element for expression of the cryparin gene. To explore the possibility of using this small fragment as a minimal core promoter, three different chimeric reporter genes were constructed using a bacterial hygromycin B resistance gene (hph), an inducible laccase of C. parasitica, and glucose oxidase of Aspergillus niger to examine the promoter properties of the fragment. When using C. parasitica as an expression host, the 188-bp fragment functioned as a promoter for the expression of all three reporter genes. Moreover, a high level of expression was further confirmed by measuring the relative amount of transcripts of hph and an internal control gene, glyceraldehyde-3-phosphate dehydrogenase, using quantitative real-time polymerase chain reaction. The 188-bp fragment also showed promoter activity in other fungi, Gibberella zeae, A. niger, and Aspergillus nidulans, which suggests that this fragment can be applied as a strong core promoter for heterologous gene expression in various fungi.
Kwon
Bo-Ra
BR
Institute for Molecular Biology and Genetics, Research Center of Bioactive Materials, Chonbuk National University, Jeonju, Chonbuk, 561-756, Korea.
Kim
Myoung-Ju
MJ
Park
Jin-A
JA
Chung
Hea-Jong
HJ
Kim
Jung-Mi
JM
Park
Seung-Moon
SM
Yun
Sung-Hwan
SH
Yang
Moon-Sik
MS
Kim
Dae-Hyuk
DH
D001483:Base Sequence; D005656:Fungal Proteins / Q000235:genetics*; D005658:Fungi / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion
C074967:CRP protein, Cryphonectria parasitica; D005656:Fungal Proteins; D008565:Membrane Proteins