Cloning directionality and terminator availability - is lac z gene terminator is enough for my gene termination?? (Jun/30/2008 )
Hello all....please help me,
I want to overexpress a plant gene by using pGreen binary vector and I am doing the cloning now. I will put the gene under its own promoter and I will insert it in SacI-XhoI (promoter to gene orientation) (please see the map of vector attached). My qustion is about terminator. Is it necessary to put my gene's natural terminator or lac Z gene got terminator? and it will do the termination job??? Hope I explained clearly. Please help me with your suggestions....thank you
-sijo-
QUOTE (sijo @ Jun 30 2008, 05:21 AM)
Hello all....please help me,
I want to overexpress a plant gene by using pGreen binary vector and I am doing the cloning now. I will put the gene under its own promoter and I will insert it in SacI-XhoI (promoter to gene orientation) (please see the map of vector attached). My qustion is about terminator. Is it necessary to put my gene's natural terminator or lac Z gene got terminator? and it will do the termination job??? Hope I explained clearly. Please help me with your suggestions....thank you
I want to overexpress a plant gene by using pGreen binary vector and I am doing the cloning now. I will put the gene under its own promoter and I will insert it in SacI-XhoI (promoter to gene orientation) (please see the map of vector attached). My qustion is about terminator. Is it necessary to put my gene's natural terminator or lac Z gene got terminator? and it will do the termination job??? Hope I explained clearly. Please help me with your suggestions....thank you
You will want to clone in the native terminator. The lac Z terminator in pgreen is not actually a terminator but rather the 3' end of the lac Z gene. This is a common arrangement used for blue-white screening to see if you have an insert (insert = lacZ reading frame lost and colony appears white).
-transient-