Difference between revisions of "PBQ200"
| (One intermediate revision by the same user not shown) | |||
| Line 8: | Line 8: | ||
*'''M13_puc_for:''' 5‘-GTAAAACGACGGCCAGTG-3‘ | *'''M13_puc_for:''' 5‘-GTAAAACGACGGCCAGTG-3‘ | ||
*'''M13_puc_rev:''' 5‘-GGAAACAGCTATGACCATG-3‘ | *'''M13_puc_rev:''' 5‘-GGAAACAGCTATGACCATG-3‘ | ||
| − | *'''FX125 (rev; primes - | + | *'''FX125 (rev; primes -85bp of MCS):''' 5‘-GGCTCGTATGTTGTGTGG-3‘ |
| + | *'''JN54 (fwd; primes +63bp of MCS):''' 5‘-GTGAAAAATGAGCCGAAAGCAG-3‘ | ||
Latest revision as of 08:12, 29 July 2014
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)
