XJb (DE3) Autolysis Glycerol Stock

T5051


XJb (DE3) Autolysis Glycerol Stock

T5051

Cat # Name Size Price Quantity
T5051 XJb (DE3) Autolysis Glycerol Stock, 1 ml 500x L-Arabinose 1 ml €109,00
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XJb (DE3) Autolysis Glycerol Stock

Highlights


  • Fast: 80 - 90% of E.coli are lysed in only 10 minutes after harvesting.
  • DE3 Lysogen: Encodes the T7 polymerase for expressing recombinant proteins under the control of the T7 promoter.
  • Versatile: Fully compatible with a wide range of buffers for protein purification and other physical methods of lysis.

Description


While there are many cell lysis methods available to scientists, unfortunately none of these methods combine all the ideal features for simple, efficient, economical, and gentle lysis of E. coli cells. The E. coli XJ autolysing strains from Zymo Research were engineered to address this problem. Mild expression of a chromosomally encoded bacteriophage λ R gene, encoding the λ lysozyme, also known as λ endolysin, is induced during growth. Cells are harvested intact while the peptidoglycan layer of the cell walls has been protected from digestion by the cytoplasmic membrane. The membrane is, however, amenable to disruption by a brief physico-chemical stress such as a freeze-thaw cycle after harvesting the cells. The XJ Autolysis™ method is highly efficient and takes only minutes (unlike traditional multiple freeze-thaw cycles). It can be applied to any number of samples without increasing processing time and labor (unlike sonication or French-press), is reliable and repeatable (unlike lysozyme treatment), and finally, is fully compatible with a wide range of buffers. Additionally, it does not require use of any potentially interfering components such as detergents, commonly found in various lytic buffers. They are also applicable for nucleic acid purification, and available with a DE3 lysogen encoding the T7 polymerase for expressing recombinant proteins driven by the T7 promoter.

Technical Specifications


Autolysis XJb lysis efficiency is 10-20 % lower than XJa. For optimal lysis, more care needs to be taken when selecting the lysis buffer. However, even very low concentrations of detergent may improve lysis significantly.
Cell Growth A very robust strain, reaching higher OD’s than E. coli K-strains.
DNA Extraction XJb is not optimal for DNA extraction.
DNA Stability This strain is RecA positive.
Genotype F- ompT hsdSB(rB - mB -) gal dcm ΔaraB::λR, cat (CmR), λ(DE3)
Processing Time 10 minutes
Product Storage -70°C to -80°C
Protein Expression XJb is ideal for recombinant protein expression. It lacks Lon and OmpT proteases, leading to higher protein yields.

Product FAQ


Q1: Is a starter culture necessary?

Q2: What buffer should the cell pellet be resuspended in?

Q3: How do you improve lysis efficiency?

Q4: What if the lysate is extremely viscous?

Q5: Can glycerol be present during the freeze-thaw cycle?

Q6: Can glucose be added to the growth media?

Q7: Will chitin be degraded?

Q8: Are competent cells GMOs?

Q9: Which is the recommended DNA concentration and volume for transformation?

Q10: Which Plasmid Size can be used for transformation?

Q11: Which antibiotics can be used with the Mix & Go! procedure?

Q12: Is it possible to dilute the competent cells?

Q13: How to reduce satellite colonies on agar plates?

Q14: How will a heat-shock affect my Transformation Efficiency?

Q15: Do the Mix & Go! strains methylate DNA?

Q16: Are the Mix & Go! strains dam+ and dcm+?

Q17: What are some tips to improve transformation efficiency?

Q18: Which strains are equivalent to the Zymo strains?

Citations


To clone new GFP-like fluorescent proteins from Obelia medusa, the authors identified the potential genes using expression libraries and cloned the genes into a vector. Expression of the proteins was facilitated by using XJb Autolysis E. coli cells from Zymo Research. The authors were able to purify three proteins from Obelia medusa that fluoresce in three different colors: cyan, green, and yellow.

Aglyamova, G.V. et al. (2011) Multi-colored homologs of the green fluorescent protein from hydromedusa Obelia sp. Photochem Photobiol Sci (8):1303-9.