发信人: else (where), 信区: Biology
标 题: Re: Kozak sequence
发信站: The unknown SPACE (Tue Jul 1 10:32:33 2003) WWW-POST

【 在 Cathleen (蛹-@-化蝶) 的大作中提到: 】
: I designed it in the primer before the FLAG tag, unfortunately after I
: sequenced the positive clone it turned out one C is missing, now the Kozak
: sequence is "CCAC" which should be "CCACC", will it still work as normal?
:
: Invitrogen really sucks, it gives two point mutations in a 58 length
: primer,fortunately, no frame shift or amino acid change, otherwise I would
: have to start all over.
:

Perfect Kozak is not as short as CCACC.
In your case, if upstream of the CACC is a G instead of A, downstream of ATG
is also a G, it maybe works (less) efficiently; otherwise, I would reconstruct
again.


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※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 130.60.]

发信人: glider (大苹果), 信区: Biology
标 题: Re: Kozak sequence
发信站: The unknown SPACE (Tue Jul 1 14:15:54 2003), 站内信件



Kozak
Sequence for
Mammalian
Expression
If you will be recombining your entry clone with a
destination vector for mammalian expression, your insert
should contain a Kozak consensus sequence with an ATG
initiation codon for proper initiation of translation (Kozak,
1987; Kozak, 1991; Kozak, 1990). An example of a Kozak
consensus sequence is provided below. Other sequences are
possible, but the G or A at position -3 and the G at position
+4 are the most critical for function (shown in bold). The
ATG initiation codon is shown underlined.
(G/A)NNATGG

【 在 Cathleen (蛹-@-化蝶) 的大作中提到: 】
: 【 在 Marble (小石头哥哥) 的大作中提到: 】
: : that is the best way. Kozac seq. needs some buffer nucleotides.
: ~~~~~~~~~~~~~~~~~~ like what?
: Kozak
: normal?
: would


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※ 来源:．The unknown SPACE bbs.mit.edu．[FROM: 128.59.]