Subject: FlyBase query
Dear Dr. Struhl,
I am curating a paper for FlyBase (Campbell and Tomlinson, 1998,
Development 125(22): 4483-4493) in which a UAS-GFP construct made by
you is used. We do not have a record for this construct in FlyBase.
I wrote to Gerard Campbell requesting some details about this construct
and he sent me the information you sent them about this line:
The UAS-GFP transgenes were in fact made by me. They're just a
GFP(S65T?)-6XMYC-NLS coding sequence stuck into the pUAST vector. You
can refer
to them as Struhl, unpublished if you like.
I am writing to check that it is OK for me to include this information
in a personal communication from you to FlyBase, so that the molecular
details of this construct make it into FlyBase.
I have a couple of questions about the construct:
a. is the GFP used the S65T version ?
b. do you know what the nuclear localisation signal (NLS) is - in the
constructs we have with NLS's, people have used 3 different sorts:
1. a 6 amino acid tag: KKKRKV
2. part of the large T antigen of SV40
3. part of the glucocorticoid receptor from rat.
I look forward to hearing from you,
Gillian
\--------------------------------------------------------------
Gillian Millburn.
FlyBase (Cambridge),
\--------------------------------------------------------------
>
Subject: Re: FlyBase query
Dear Gillian,
In answer to your questions:
a - it is the S65T version of GFP
b - it is the SV40 NLS PPKKKRKVED
The same coding sequence was used in a hs-GFP-MYC-NLS transgene published in
Jiang and Struhl, Nature, 1998.
Hope that helps.
Best wishes.
Gary
> Dear Dr. Struhl,
>
> I am curating a paper for FlyBase (Campbell and Tomlinson, 1998,
> Development 125(22): 4483-4493) in which a UAS-GFP construct made by
> you is used. We do not have a record for this construct in FlyBase.
>
> I wrote to Gerard Campbell requesting some details about this construct
> and he sent me the information you sent them about this line:
>
> The UAS-GFP transgenes were in fact made by me. They're just a
> GFP(S65T?)-6XMYC-NLS coding sequence stuck into the pUAST vector. You
> can refer
> to them as Struhl, unpublished if you like.
>
> I am writing to check that it is OK for me to include this information
> in a personal communication from you to FlyBase, so that the molecular
> details of this construct make it into FlyBase.
>
> I have a couple of questions about the construct:
>
> a. is the GFP used the S65T version ?
> b. do you know what the nuclear localisation signal (NLS) is - in the
> constructs we have with NLS's, people have used 3 different sorts:
>
> 1. a 6 amino acid tag: KKKRKV
>
> 2. part of the large T antigen of SV40
>
> 3. part of the glucocorticoid receptor from rat.
>
> I look forward to hearing from you,
>
> Gillian
>
> --------------------------------------------------------------
> Gillian Millburn.
>
> FlyBase (Cambridge),
> --------------------------------------------------------------