pSecTag2 A, B, & C Mammalian Expression Vectors
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Citations & References (20)
Invitrogen™

pSecTag2 A, B, & C Mammalian Expression Vectors

pSecTag2およびpSecTag2/Hygroは、融合タンパク質の分泌、浄化、および検出用に設計された哺乳類発現ベクターです。各ベクターには、3つの測定フレーム内に大規模な複数のクローニング部位があるため、N末端分泌シグナルによるフレーム内でのクローニングが簡素化されます。ベクター（図1）には以下の特長があります詳細を見る
テクニカルサポートオーダーサポート
製品番号（カタログ番号）数量
V9002020 μg
製品番号（カタログ番号） V90020
価格（JPY）
147,900
20 µg
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数量:
20 μg
pSecTag2およびpSecTag2/Hygroは、融合タンパク質の分泌、浄化、および検出用に設計された哺乳類発現ベクターです。各ベクターには、3つの測定フレーム内に大規模な複数のクローニング部位があるため、N末端分泌シグナルによるフレーム内でのクローニングが簡素化されます。ベクター（図1）には以下の特長があります。

•組換えタンパク質の効率的な分泌のためのマウスIgカッパ鎖のV-J2-C領域からの分泌シグナル（図2）
• 高レベルで構成的な発現のためのサイトメガロウイルス（CMV）プロモーター
• ニッケルキレート樹脂による迅速な浄化用のC末端ポリヒスチジン（6xHis）タグと抗His（C-term）抗体による検出
• 抗myc抗体による検出および分析用のC末端c-mycエピトープ
•ウシ成長ホルモン（BGH）ポリアデニル化シグナルおよびmRNA安定性向上のための転写終了配列
• ラージT抗原（COS-1やCOS-7など）を発現する細胞株内でのエピソーム複製および単純なベクターレスキュー用のSV40起源

pSecTag2ベクターは、哺乳類細胞の費用対効果の高い選択のために、ゼオシン™耐性遺伝子を持っています。ゼオシン™の選択は、大腸菌でも使用できます。

pSecTag2/Hygroベクターには、安定した哺乳類細胞株を選択するためのハイグロマイシンB耐性遺伝子があります。
研究用にのみ使用できます。診断用には使用いただけません。
仕様
供給タイプTransfection
使用対象（アプリケーション）分泌発現
製品タイプ哺乳類発現用ベクター
数量20 μg
選択剤（真核生物）ゼオシン™
ベクターpSec
クローニング法制限酵素／MCS
プロモーターCMV
タンパク質タグHisタグ（6x）、c-Mycエピトープタグ、IgKリーダー配列, c-Myc Epitope Tag, IgK Leader Sequence
Unit Size20 µg
組成および保存条件
それぞれ20 µgのベクターとPSA発現コントロールが超らせん型で凍結乾燥された状態で提供されます。-20℃で保存ベクターは、適切に保存されている場合、6カ月間安定していることが保証されます。

よくあるご質問（FAQ）

I used the pSecTag2 vector and am seeing cellular but not secreted expression of the protein of interest. Can you offer some tips?

If the gene of interest has a start codon in the context of a perfect Kozak sequence, it may be preferentially translated over the vector’'s initiation codon, resulting in no leader sequence and no glycosylation, and hence no targeting to the endoplasmic reticulum and no secretion. This is rare, but it has been observed. If it is a problem, we recommend using PCR to delete the start codon.

Can I propagate my pSecTag2 vector in E. coli containing the Tn5 gene?

pSectag2 vectors have the Zeocin antibiotic-resistance marker for selection in E. coli, and any E. coli strain that contains the complete Tn5 transposable element (i.e., DH5alphaF'IQ, SURE, SURE2) encodes the ble (bleomycin) resistance gene that confers resistance to the Zeocin antibiotic. Hence, for the most efficient selection, we highly recommend choosing an E. coli strain that does not contain the Tn5 gene.

I see that the pSectag2 vectors carry the Zeocin antibiotic-resistance gene driven by the EM7 promoter. Can I use Zeocin antibiotic for selection when I propagate these vectors in E. coli?

Yes, you can use Zeocin antibiotic for selection in E. coli. However, keep in mind that for Zeocin antibiotic to be active, the salt concentration of the medium must remain low (<90 mM) and the pH must be 7.5. Prepare LB broth and LB agar plates using low-salt (5 g NaCl/liter) LB.

What is the difference between pSecTag2 and pSecTag2/Hygro vectors?

The only difference between these vectors is that the pSecTag2 vectors have the Zeocin antibiotic-resistance gene for stable selection, whereas the pSecTag2/Hygro vectors have the Hygromycin B resistance gene for stable selection.

I would like to get secreted expression of my protein. What are your recommendations?

You can insert an N-terminal secretion signal or leader sequence upstream of your gene and in-frame with the gene sequence to facilitate secreted expression of the protein. We actually offer the pSecTag2 (Cat. No. V90020) and pSecTag2/Hygro (Cat. No. V91020) vectors designed for this purpose. These vectors contain the N-terminal murine Ig kappa-chain secretion signal for secreted expression of the protein of interest.

View all pSecTag2 A, B, & C Mammalian Expression Vectors FAQs

引用および参考文献 (20)

引用および参考文献
Abstract
Neuropilin-1 binds vascular endothelial growth factor 165, placenta growth factor-2, and heparin via its b1b2 domain.
Authors: Mamluk Roni; Gechtman Ze'ev; Kutcher Matthew E; Gasiunas Nijole; Gallagher John; Klagsbrun Michael;
Journal:J Biol Chem
PubMed ID:11986311
'Neuroplin-1 (NRP1), a receptor for vascular endothelial growth factor (VEGF) family members, has three distinct extracellular domains, a1a2, b1b2, and c. To determine the VEGF(165) and placenta growth factor 2 (PlGF-2)-binding sites of NRP1, recombinant NRP1 domains were expressed in mammalian cells as Myc-tagged, soluble proteins, and used in co-precipitation ... More
Molecular dissection of the alpha-dystroglycan- and integrin-binding sites within the globular domain of human laminin-10.
Authors:Ido H, Harada K, Futaki S, Hayashi Y, Nishiuchi R, Natsuka Y, Li S, Wada Y, Combs AC, Ervasti JM, Sekiguchi K,
Journal:J Biol Chem
PubMed ID:14701821
'The adhesive interactions of cells with laminins are mediated by integrins and non-integrin-type receptors such as alpha-dystroglycan and syndecans. Laminins bind to these receptors at the C-terminal globular domain of their alpha chains, but the regions recognized by these receptors have not been mapped precisely. In this study, we sought ... More
Sulfation of N-acetylglucosamine by chondroitin 6-sulfotransferase 2 (GST-5).
Authors: Bhakta S; Bartes A; Bowman K G; Kao W M; Polsky I; Lee J K; Cook B N; Bruehl R E; Rosen S D; Bertozzi C R; Hemmerich S;
Journal:J Biol Chem
PubMed ID:10956661
'Based on sequence homology with a previously cloned human GlcNAc 6-O-sulfotransferase, we have identified an open reading frame (ORF) encoding a novel member of the Gal/GalNAc/GlcNAc 6-O-sulfotransferase (GST) family termed GST-5 on the human X chromosome (band Xp11). GST-5 has recently been characterized as a novel GalNAc 6-O-sulfotransferase termed chondroitin ... More
Plexin-B1 directly interacts with PDZ-RhoGEF/LARG to regulate RhoA and growth cone morphology.
Authors: Swiercz Jakub M; Kuner Rohini; Behrens Jürgen; Offermanns Stefan;
Journal:Neuron
PubMed ID:12123608
'Plexins are widely expressed transmembrane proteins that, in the nervous system, mediate repulsive signals of semaphorins. However, the molecular nature of plexin-mediated signal transduction remains poorly understood. Here, we demonstrate that plexin-B family members associate through their C termini with the Rho guanine nucleotide exchange factors PDZ-RhoGEF and LARG. Activation ... More
Death receptor 5, a new member of the TNFR family, and DR4 induce FADD- dependent apoptosis and activate the NF-kappaB pathway.
Authors:Chaudhary PM, Eby M, Jasmin A, Bookwalter A, Murray J, Hood L
Journal:Immunity
PubMed ID:9430227
'Death receptor 4 (DR4) is a recently described receptor for the cytotoxic ligand TRAIL that reportedly uses a FADD-independent pathway to induce apoptosis and does not activate the NF-kappaB pathway. We have isolated a new member of the tumor necrosis factor receptor (TNFR) family, designated DR5, which bears a high ... More
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