Invitrogen™ One Shot™ BL21(DE3)pLysS Chemically CompetentE. coli

Description

• One Shot BL21(DE3)pLysS Chemically Competent E
• coli cells are ideal for use with bacteriophage T7 promoter-based expression systems such as pRSET and pET
• The pLysS plasmid carried by the BL21(DE3) pLysS strain produces T7 lysozyme, an inhibitor of T7 RNA polymerase, to reduce basal level expression of the gene of interest
• pLysS confers resistance to chloramphenicol (CamR) and contains the p15A origin
• This origin allows pLysS to be compatible with plasmids containing the ColE1 or pMB1 origin (i.e
• pUC- or pBR322- derived plasmids)
• Recombinant proteins that are nontoxic to E
• coli are generally expressed at higher levels in BL21(DE3) cells compared to the BL21(DE3) pLysS strain with tighter expression control
• BL21(DE3)pLysS is recommended for use when expressing toxic genes
• The BL21 (DE3) E
• coli strain is one of the most popular host strains to produce recombinant proteins
• This strain had been engineered to carry the λDE3 lysogen that contains T7 RNA polymerase gene under control of the lac UV5 promoter
• IPTG is required to induce expression of the T7 RNA polymerase cascade system that in turn transcribe T7 promoter regulated target gene
• BL21(DE3) is a derivative from the E
• coli B strain that does not contain the ion protease and is also deficient in the outer membrane protease OmpT
• The lack of two key proteases reduces degradation of heterologous proteins expressed in the strains
• Additionally, several other strain features make them especially suited for protein production, namely fast growth in minimal medium, a lower acetate production when grown in high glucose media, and an ability to reach high cell density
• One Shot BL21(DE3)pLysS Chemically Competent E
• coli cells can achieve transformation efficiency of >1 x 10 8 cfu/μg of control DNA
• One Shot BL21(DE3)pLysS Chemically Competent E
• coli cells offer: • Transformation efficiency of >1 x 10 8 cfu/μg control plasmid DNA • Ion and OmpT protease deficiency that reduces degradation of recombinant protein • IPTG-inducible lac UV5 promoter that controls expression of the T7 RNA polymerase • pLysS (CamR) plasmid for reduced basal expression in uninduced cells • hsd S mutation that allows efficient transformation of unmethylated DNA • Fast growth in minimal medium and ability to reach high cell density Easy-to-use One Shot format BL21(DE3)pLysS Chemically Competent E
• coli cells are supplied in the convenient, single-reaction One Shot format
• The single-tube, single-use format allows all steps of the transformation protocol, up to plating, to take place in the same tube, thereby helping save time and to prevent contamination
• Genotype F – omp T hsd SB (r B –, m B –) gal dcm (DE3) pLysS(CamR) Find the strain and format that you need • We offer other E
• coli strains for protein expression
• • For expression of toxic proteins, consider BL21 AI One Shot Chemically Competent E
• coli (Cat
• No
• C607003)
• • BL21 Star (DE3) and other strains are available in MultiShot formats for high throughput applications
• • We offer a variety of systems for the expression of recombinant proteins in E
• coli
• The Champion pET expression system provides the highest level of protein production available in any expression system.