Hi Towa,
The LacZ gene can be broken in two expression units - LacZ-alpha and LacZ-omega. The pBluescript (and pUC series) plasmids, as Koeng pointed out, has the much smaller LacZ-alpha gene. LacZ-alpha by itself will not give you beta-gal activity. If you use this then you will need a bacterial host (e.g. DH5-alpha) that contains the omega gene fragment - lacZΔM15. The combination of the alpha and omega gene products produces functional beta-gal activity in what is call alpha-complementation. So it depends on what you want to do in your experiment as to which plasmid to use. Full length lacZ will allow you to express beta-gal in almost any cell type but if you want to do blue/white selection then you will need alpha-complementation with the appropriate plasmid and host.Cheers,
Scott
http://www.opensciencenet.org
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diybio@googlegroups.com. To unsubscribe from this group, send email to diybio+unsubscribe@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+unsubscribe@googlegroups.com.
To post to this group, send email to diybio@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/f5cde38b-5951-4ff4-a05f-e64329d4f803%40googlegroups.com.
For more options, visit https://groups.google.com/d/optout.






0 comments:
Post a Comment