Analysis of the VPE sequences in the Caenorhabditis elegans vit-2 promoter with extrachromosomal tandem array-containing transgenic strains
| Autor: | John Spieth, Thomas Blumenthal, C Madej, Kristi Lea, Margaret MacMorris |
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| Rok vydání: | 1994 |
| Předmět: |
Caenorhabditis briggsae
Molecular Sequence Data Mutant Biology Animals Genetically Modified Fusion gene Vitellogenins Transformation Genetic Species Specificity Extrachromosomal DNA Animals Caenorhabditis elegans Promoter Regions Genetic Gene Molecular Biology Genes Helminth Sequence Deletion Genetics Regulation of gene expression Base Sequence Genetic Variation DNA Cell Biology biology.organism_classification Gene Expression Regulation Regulatory sequence Research Article |
| Zdroj: | Molecular and Cellular Biology. 14:484-491 |
| ISSN: | 1098-5549 |
| DOI: | 10.1128/mcb.14.1.484-491.1994 |
| Popis: | The Caenorhabditis elegans vit genes, encoding vitellogenins, are abundantly expressed in the adult hermaphrodite intestine. Two repeated elements, vit promoter element 1 (VPE1 [TGTCAAT]) and VPE2 (CTGATAA), have been identified in the 5' flanking DNA of each of the vit genes of C. elegans and Caenorhabditis briggsae. These elements have previously been shown to be needed for correctly regulated expression of a vit-2/vit-6 fusion gene in low-copy-number, integrated transgenes. Here we extend the analysis of the function of VPE1 and VPE2 by using transgenic lines carrying large, extrachromosomal arrays of the test genes. The results validate the use of such arrays for transgenic analysis of gene regulation in C. elegans, by confirming previous findings showing that the VPE1 at -45 and both VPE2s are sites of activation. Additional experiments now indicate that when the -45 VPE1 is inverted or replaced by a VPE2, nearly total loss of promoter function results, suggesting that the highly conserved -45 VPE1 plays a unique role in vit-2 promoter function. In contrast, single mutations eliminating the three upstream VPE1s are without effect. However, in combination in double and triple mutants, these upstream VPE1 mutations cause drastic reductions in expression levels. The -150 VPE2 can be replaced by a XhoI site (CTCGAG), and the -90 VPE2 can be eliminated, as long as the overlapping VPE1 is left intact, but when these two replacements are combined, activity is lost. Thus, the promoter must have at least one VPE2 and it must have at least two VPE1s, one at -45 and one additional upstream element. |
| Databáze: | OpenAIRE |
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