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ハエの培養細胞に使えそうな発現ベクター


Date: Tue, 13 Oct 1998 08:37:47 +0900
From: tmurataアットマークrtc.riken.go.jp (MuraTa, Riken Tsukuba)
Subject: [Jfly] DES vectors.

村田アットマーク理研です。
以前、ハエの培養細胞への遺伝子導入&発現の質問が出ていましたが、それに関連して。
いくつかの会社に発現ベクターについて質問したところ、まずは以下のような返事が
来ました。
参考にしてください。

Dear Dr. Murata,

Thank you for your interest in Invitrogen's Drosophila Expression System
(DES). There are several vectors available for Invitrogen's DES system.
There are three general vectors available at the initial purchase of a DES
kit. They are pAc5.1/V5-His, pMT/V5-His, and the pMT/BiP/V5-His vectors.

pAc5.1/V5-His:
This vector contains a strong Actin 5C promoter for high-level constitutive
expression of your protein. It contains the V5 epitope and polyhistidine
tag at the C-terminus of the multiple cloning site (MCS) to aid you in
detecting your protein with the anti-V5 antibody (Cat. No. R960-25) and
isolating your protein via a nickle chelated resin such as our Probond
resin (Cat. No. R801-01).

pMT/V5-HIs:
This vector uses the Drosophila metallothionein promoter for high-level
inducible expression of your protein upon addition of copper sulfate or
calcium chloride to the medium. Once again, this vector has the V5 epitope
and polyhistidine tag at the C-terminus of the MCS.

pMT/BiP/V5-His:
This vectors uses the Drosophila metallothionein promoter for high-level
inducible expression of your protein upon addition of copper sulfate or
calcium chloride to the medium. It also adds an N-terminal secretion
signal to your protein to direct it to the culture medium. This ensures
proper posttranslational modifications and facilitates purification of your
protein. Once again, this vector has the V5 epitope and polyhistidine tag
at the C-terminus of the MCS.

Invitrogen carries one other DES vector, pDS47/V5-His. However, this
vector can only be purchased once a complete DES kit has been purchased.
This particular vector is not provided in kit format. The pDS47/V5-HIs
contains a moderately strong constitutive vector for use with DES. Once
again, this vector has the V5 epitope and polyhistidine tag at the
C-terminus of the MCS.

All of the above mentioned vectors is provided with the MCS in three
reading frames relative to the C-terminal tag to facilitate cloning of your
gene of interest.

Please note, in order to purchase a DES kit, you must sign a DES license
agreement. The license agreement can be downloaded from the following web
address:

http://www.invitrogen.com/dros/des_license.html

Once you are ready to purchase the DES kit, please contact our Japanese
distributor, Funakoshi Co. Ltd. at the following contact address:

Funakoshi Co. Ltd.
9-7 Hongo 2-Chome
Bunkyo-ku, Tokyo 113
Phone: 03-5684-1616
Fax: 03-5684-1633
E-mail: funaアットマークgol.com
Web Site: http://www.funakoshi.co.jp

Sincerely,
Hiromi Kimura
International Technical Specialist
Invitrogen Corporation


村田 武英
305-0074 茨城県 つくば市 高野台 3-1-1
理化学研究所 ライフサイエンス筑波研究センター