Difference between revisions of "Part:BBa K3970002"

(Usage and Biology)
 
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The BS module，which is the lexO is the diversification and characterization of the LexA-binding motifs. We find it in an paper.
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__NOTOC__
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The previously characterized LexA binding motif has a consensus sequence of a 16 bp palindrome, CTGTATATATATACAG. Yeast contains a number of genes regulated by the LexA repressor. These genes, as well as the previously used modified GAL1 promoter with 8 LexA BS, contain a spectrum of sequence modifications deviating from the consensus LexA binding site .Based on the native LexA binding sites present in the regulatory region of the lexA gene and in the ColEI operator, the author designed versions of the LexA BS with minor sequence modifications and tested their binding affinity to purified sTF16 in the electrophoretic mobility shift assay (EMSA) . The sTF16 used was composed of a LexA DNA binding domain, the Herpes simplex virus transactivation domain (VP16) and a 6×His-tag. The results indicate that all four versions of the BSs bind similarly to the LexA-DNA-binding domain in vitro and therefore can also be expected to perform similarly within a synthetic output promoter in vivo.
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<partinfo>BBa_K3970002 short</partinfo>
  
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Reference:
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==Usage and Biology==
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Synthetic Transcription Amplifier System for Orthogonal Control of Gene Expression in Saccharomyces cerevisiae;PLOS ONE | DOI:10.1371/journal.pone.0148320
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URA3 gene is a gene on chromosome V of yeast, whose systematic name is YEL021W. It encodes the whyside 5-phosphate decarboxylase, an enzyme that catalyzes one of the key reactions in the synthesis of pyrimidine nucleotides from yeast RNA.  The whyside 5-phosphate decarboxylase is inactivated and the yeast cannot grow unless uridine or uracil is added to the medium.  If you were to nutrient transformation into URA3 gene defects strains, as these nutritional deficiencies plants can grow (positive), on the contrary, if you add 5- to medium FOA (5-fluorine orotic acid), then the original keep normal type yeast cells of whey 5-phosphoric acid decarboxylase can convert 5-FOA into toxic substances, (suicide inhibitor) and its product,  5-fluorouracil can cause cell death (negative selection).
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Because URA3 gene can carry out both positive and negative selection, it has become a genetic marker, which is widely used in gene transformation and other genetic experiments of bacteria and many kinds of fungi.  It is also one of the most important genetic markers (reporter genes) in yeast genetic engineering.
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3970002 SequenceAndFeatures</partinfo>
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K3970002 parameters</partinfo>
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Latest revision as of 12:07, 8 October 2021

URA3

Usage and Biology

URA3 gene is a gene on chromosome V of yeast, whose systematic name is YEL021W. It encodes the whyside 5-phosphate decarboxylase, an enzyme that catalyzes one of the key reactions in the synthesis of pyrimidine nucleotides from yeast RNA.  The whyside 5-phosphate decarboxylase is inactivated and the yeast cannot grow unless uridine or uracil is added to the medium.  If you were to nutrient transformation into URA3 gene defects strains, as these nutritional deficiencies plants can grow (positive), on the contrary, if you add 5- to medium FOA (5-fluorine orotic acid), then the original keep normal type yeast cells of whey 5-phosphoric acid decarboxylase can convert 5-FOA into toxic substances, (suicide inhibitor) and its product,  5-fluorouracil can cause cell death (negative selection).

Because URA3 gene can carry out both positive and negative selection, it has become a genetic marker, which is widely used in gene transformation and other genetic experiments of bacteria and many kinds of fungi.  It is also one of the most important genetic markers (reporter genes) in yeast genetic engineering.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]