What are the examples? Is there any reading material available on this? I googled, but could not find proper information.
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Ram
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#1
ram
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Posted 11 June 2009 - 05:21 AM
What is stringent vector? Is the term 'stringent' related to level of expression? What are other types of expression vectors?
What are the examples? Is there any reading material available on this? I googled, but could not find proper information.
Thanks
Ram
What are the examples? Is there any reading material available on this? I googled, but could not find proper information.
Thanks
Ram
If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.
#2
genz
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Posted 11 June 2009 - 04:45 PM
Assume you express your protein in E coli.
There are many vectors, some use T7 promoter, some lac, T7/lac etc... Each promoter has different stringency. T7 is known to be leaky, so some vector will include a lacI to prevent the leakage, some will have pLysS etc.
try http://www.bio.net/b...rch/074525.html
There are many vectors, some use T7 promoter, some lac, T7/lac etc... Each promoter has different stringency. T7 is known to be leaky, so some vector will include a lacI to prevent the leakage, some will have pLysS etc.
try http://www.bio.net/b...rch/074525.html
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ram
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Posted 12 June 2009 - 01:12 AM
Thank you for the information.
Yes. Its for expression in E coli
So the adjective 'stringent' before vector is always related to the promoter of the vector..right?
As you said, expression systems consisting of T7 promoter are considered to be leaky, meaning they are not stringent. Why pET vectors (containing T7 promoter) are considered stringent? Can you comment anything on 'stringency' of tet promoter systems (eg. pASK-IBA7) ? If I am getting very low level of expression with pASK-IBA7 vector, does it mean that I may get good levels in pET since this is not a stringent vector (or is it?) CONFUSED! What are other non-stringent vectors?
The link that u posted is not opening
Yes. Its for expression in E coli
So the adjective 'stringent' before vector is always related to the promoter of the vector..right?
As you said, expression systems consisting of T7 promoter are considered to be leaky, meaning they are not stringent. Why pET vectors (containing T7 promoter) are considered stringent? Can you comment anything on 'stringency' of tet promoter systems (eg. pASK-IBA7) ? If I am getting very low level of expression with pASK-IBA7 vector, does it mean that I may get good levels in pET since this is not a stringent vector (or is it?) CONFUSED! What are other non-stringent vectors?
The link that u posted is not opening
If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.
