Upon induction with IPTG in TB media, cells turned into pinkish color and yielded brownish inclusion body; while same BL21 culture inoculated in LB media resulting in green color.
I have been tested for iron in green color LB media, it showed aprx. 0.2ppm of iron. But still I am doubtful.
Facts:
-Recombinant protein is 6.7kDa, rich in beta sheets.
-No contamination in culture and kanamycin has been used in both cultures.
Hi,
Does your protein bind the metal ions? Your results suggests your protein binds some metal ions if the media isn't polluted by other bacteria.
Sir,
I have performed gram staining to confirm the purity of culture (image attached). I have no idea about its metal binding property from its available literature. Is there any way by which I can find out the metal binding property of the protein?
Hai vikas, Vikas K Singh did you get any answers why your BL21 DE3 turns the medium green?
I've seen this before in some cultures at the Fac with these various reasons; I'd try to check if the following might be the case:
1) if TB is buffering correctly, and if your LB is not buffered; try the modified recipe that has added buffering
2) if enough antibiotics or TOO much has been added, if the flasks have been closed cleaning, sometimes contamination with other aerobic/anaerobic strains resistant to your resistance gene can occur leading to fermentation of the media with colored appearances
3) if the TB/LB recipes are correct and if the purity of the LB is up to scratch. Most Tryptone/peptone powders are not identical even from batch to batch, lot to lot or even company to company
4) Powders of lower quality have high trace metal levels, and the media powders aren't standardized; if all else fails order premixed lb media mix
I've seen this before in some cultures at the Fac with these various reasons; I'd try to check if the following might be the case:
1) if TB is buffering correctly, and if your LB is not buffered; try the modified recipe that has added buffering
2) if enough antibiotics or TOO much has been added, if the flasks have been closed cleaning, sometimes contamination with other aerobic/anaerobic strains resistant to your resistance gene can occur leading to fermentation of the media with colored appearances
3) if the TB/LB recipes are correct and if the purity of the LB is up to scratch. Most Tryptone/peptone powders are not identical even from batch to batch, lot to lot or even company to company
4) Powders of lower quality have high trace metal levels, and the media powders aren't standardized; if all else fails order premixed lb media mix
- Badges
- Science method