Difference between revisions of "PBQ200"

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(New page: vector for the overexpression of proteins in ''B. subtilis'' constructed in the G. Rapoport lab in ''E. coli'': ampicillin resistance in ''B. subtilis'': erythromycin resistance based o...)
 
 
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vector for the overexpression of proteins in ''B. subtilis''
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[[File:PBQ200.jpeg|250px|thumb|right|'''pBQ200: click to enlarge''']]
  
constructed in the G. Rapoport lab
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'''Application:'''
  
in ''E. coli'': ampicillin resistance
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The vector was constructed in the George Rapoport lab and it is suitable for constitutive overexpression of proteins in ''B. subtilis''. The plasmid confers resistance to ampicillin and erythromycin in ''E. coli'' and ''B. subtilis'', respectively. [[pBQ200]] is based on the vector pHT315. A Shine-Dalgarno sequence has to be fused to the open reading frame during PCR.
in ''B. subtilis'': erythromycin resistance
 
  
based on [[pHT315]]
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Sequencing primers:
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*'''M13_puc_for:'''                      5‘-GTAAAACGACGGCCAGTG-3‘
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*'''M13_puc_rev:'''                      5‘-GGAAACAGCTATGACCATG-3‘
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*'''FX125 (rev; primes -85bp of MCS):''' 5‘-GGCTCGTATGTTGTGTGG-3‘
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*'''JN54 (fwd; primes +63bp of MCS):''' 5‘-GTGAAAAATGAGCCGAAAGCAG-3‘
  
Martin-Verstraete et al. (194) Interactions of wild type and truncated LevR of Bacillus subtilis with the upstream activating sequence of the levanase operon. J. Mol. Biol. 241: 178-192. [http://www.ncbi.nlm.nih.gov/pubmed/8057358?ordinalpos=5&itool=EntrezSystem2.PEntrez.Pubmed.Pubmed_ResultsPanel.Pubmed_DefaultReportPanel.Pubmed_RVDocSum PubMed]
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'''Reference:'''
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<pubmed> 8057358 </pubmed>
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== Return to [[Plasmids]] ==

Latest revision as of 08:12, 29 July 2014

pBQ200: click to enlarge

Application:

The vector was constructed in the George Rapoport lab and it is suitable for constitutive overexpression of proteins in B. subtilis. The plasmid confers resistance to ampicillin and erythromycin in E. coli and B. subtilis, respectively. pBQ200 is based on the vector pHT315. A Shine-Dalgarno sequence has to be fused to the open reading frame during PCR.

Sequencing primers:

  • M13_puc_for: 5‘-GTAAAACGACGGCCAGTG-3‘
  • M13_puc_rev: 5‘-GGAAACAGCTATGACCATG-3‘
  • FX125 (rev; primes -85bp of MCS): 5‘-GGCTCGTATGTTGTGTGG-3‘
  • JN54 (fwd; primes +63bp of MCS): 5‘-GTGAAAAATGAGCCGAAAGCAG-3‘


Reference:

I Martin-Verstraete, M Débarbouillé, A Klier, G Rapoport
Interactions of wild-type and truncated LevR of Bacillus subtilis with the upstream activating sequence of the levanase operon.
J Mol Biol: 1994, 241(2);178-92
[PubMed:8057358] [WorldCat.org] [DOI] (P p)

Return to Plasmids