[Arabidopsis] Questions regarding a big gene

George E. Schaller george.e.schaller at Dartmouth.EDU
Thu Apr 20 10:59:06 EST 2006


I would not believe in the subcellular localization unless it  
included the entire coding sequence. Otherwise you may have deleted  
an important motif. Additionally, if you use a partial sequence it  
may not fold correctly--which could lead to all sorts of alternative  
spurious localizations.--E

On Apr 19, 2006, at 11:47 AM, wang zhibib wrote:

> Hi Everyone,
>
> I am wondering if anyone could help me the following
> questions.  I am working on a big gene more than 10kb.
>
> 1. I had problem with RNA blotting. Any tip to
> transfer total RNA well to filter;
> 2. Protein sequence analyses identified three NLS
> (nuclear localization signal) and dispersed in gene.
> Do I need absolutely include three NLS to determine
> its subcellular localization? It took too much time
> for me to get the full length cDNA. Will it be
> sufficient to use one or two NLS to fuse GFP?
>
> Thank you in advance.
>
> Zhibin
>
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G. Eric Schaller
Dept. of Biological Sciences
Dartmouth College
Hanover, NH 03755
Phn: 603-646-2525
Fax: 603-646-1347






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