Subject: FlyBase query
Dear Dr. Marti-Subirana,
I am curating a paper for FlyBase (Buescher and Chia, 1997, Development 124
(3) 673-681) in which a TaulacZ cassette obtained from you and R. Holmgren
is used.
I would be really grateful if you could let me know the details of how the
cassette is constructed, for example,
1) what promoter drives expression of the taulacZ fusion gene?
2) what is the nature of the taulacZ fusion gene - is the tau gene used
bovine tau, and which amino acids/domains from tau and lacZ are present?
3) what is the marker gene in the FRT cassette?
4) where is the FRT cassette located within the taulacZ gene - is it
between the promoter and the taulacZ coding region?
Also, is there a paper in which these details are published?
As this construct is new to FlyBase I would like to get some data from you
and/or R. Holmgren, and not just a secondary publication.
I look forward to hearing from you,
Gillian Millburn
\--------------------------------------------------------------
Gillian Millburn.
FlyBase (Cambridge),
\--------------------------------------------------------------
>
Subject: Re: FlyBase query
Dear Gillian Millburn,
	The 'flip-out construct is described in Buenzow and Holmgren (1995)
Developmental biology  170:338-349 .
	Our paper using this construct is being resubmitted for publication.
	1) promoter: actin5C
	2) The tau-lacZ is from Callahan and Thomas (1994) PNAS 91: 5972-5976.
	3) There is no marker in the 'flip-out cassette', though there is a
Not 1 cloning site for that purpose.
	4) The 'flip-out' cassette is between the promoter and the coding
sequences for tau-lacZ.
\--
Robert Holmgren
Northwestern University
>
Subject: Re: FlyBase query
Dear Dr. Millburn,
The fragment containing the in-frame fusion between the bovine tau cDNA and=
the
lacZ gene was obtained from Callahan and Thomas (PNAS 91: 5972-5976, 1994) =
and
cloned into the unique SalI site of the Carnegie20/Actin5C/FRT cassete vect=
or
(Buenzow and Holmgren, 1995, Dev. Biol.  170:338-349 ).
To answer your specific questions:
1) the actin 5C promoter
2) refer to Callahan and Thomas (1994)
3) the A5C/FC/taulacZ is marked with ry+
4) the FRT sites are located within the promoter and the reporter taulacZ g=
ene
You will find details of both in the references mentioned above.
We have recently submitted a paper in which the construct is detailed and t=
hat
would be the reference for the construct. I guess it has to be referred to=
as
=22unpublished result=22 until it is published. I could send you the refere=
nce for
the paper when it is accepted for publication.
Please let me know if I can be of further help,
Anna Marti-Subirana
___________________________________________________________________________=
____
Subject: FlyBase query
Dear Dr. Marti-Subirana,
I am curating a paper for FlyBase (Buescher and Chia, 1997, Development 124
(3) 673-681) in which a TaulacZ cassette obtained from you and R. Holmgren
is used.
I would be really grateful if you could let me know the details of how the
cassette is constructed, for example,
1) what promoter drives expression of the taulacZ fusion gene?
2) what is the nature of the taulacZ fusion gene - is the tau gene used
bovine tau, and which amino acids/domains from tau and lacZ are present?
3) what is the marker gene in the FRT cassette?
4) where is the FRT cassette located within the taulacZ gene - is it
between the promoter and the taulacZ coding region?
Also, is there a paper in which these details are published?
As this construct is new to FlyBase I would like to get some data from you
and/or R. Holmgren, and not just a secondary publication.
I look forward to hearing from you,
Gillian Millburn
\--------------------------------------------------------------
Gillian Millburn.
FlyBase (Cambridge),
\--------------------------------------------------------------