pYES2.1 TOPO™ TA Yeast Expression Kit

Citations & References (7)

Invitrogen™

pYES2.1 TOPO™ TA Yeast Expression Kit

pYES2.1 TOPO TA Expression Kitでは、Taq増幅PCR産物をSaccharomyces cerevisiae発現ベクターに直接クローニングできます。このキットでは、直線化されたトポイソメラーゼI活性化pYES2.1/V5-His-TOPO¤ベクターをベンチトップで5分間のクローニングに使用して、85詳細を見る

製品番号（カタログ番号）	数量
K415001	20 reactions

製品番号（カタログ番号） K415001

価格（JPY）

94,700

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Ends: 26-Dec-2025

157,900

割引額 63,200 (40%)

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20 reactions

pYES2.1 TOPO TA Expression Kitでは、Taq増幅PCR産物をSaccharomyces cerevisiae発現ベクターに直接クローニングできます。このキットでは、直線化されたトポイソメラーゼI活性化pYES2.1/V5-His-TOPO¤ベクターをベンチトップで5分間のクローニングに使用して、85 %の組換えを行います。pYES2.1/V2- His-TOPO™ベクターの特長：

•経済的な最小限の培地での栄養要求性の選択用の遺伝子の高コピー維持のための2µの複製元
• 効率的な検出および浄化のためのC末端V5エピトープおよびポリヒスチジン（6xHis）タグ

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。

抗生物質耐性菌アンピシリン（AmpR）

製品タイプTA Yeast Expression Kit

数量20 reactions

ベクターpYES、TOPO-TAクローニングベクター

クローニング法TOPO™-TA

製品ラインTOPO、あり, YES

プロモーターGAL1

タンパク質タグHisタグ（6x）、V5エピトープタグ, V5 Epitope Tag

Unit SizeEach

組成および保存条件

各pYES2.1 TOPO™ TA Expression Kitには2つのボックスが含まれています。pYES2.1 TOPO™ TAボックスには、200 ngのトポイソメラーゼI活性化pYES2.1/V5-His-TOPO™ベクター、滅菌水、dNTP、10X PCRバッファー、塩溶液、コントロールテンプレートおよびプライマー、シーケンシングまたはPCRスクリーニングのためのGAL1フォワードプライマーおよびV5 C-termリバースプライマー、および発現コントロールプラスミドなど、ライゲーションに必要なすべての試薬が含まれています。-20℃で保存One Shot™ Boxボックスには、ケミカルコンピテントTOP10Fの大腸菌の50µLのアリコート21個、S.O.C.培地、超らせん型プラスミドコントロールをpUC19含めて、形質転換に必要なすべての試薬が含まれています。-80℃で保存してください。すべての成分は、指定された温度で適切に保存されている場合、6ケ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）

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

What are the different kinds of media used for culturing Pichia pastoris and S. cerevisiae?

Following are the rich and minimal media used for culturing Pichia pastoris and S. cerevisiae:

Rich Media:
S. cerevisiae and Pichia pastoris
YPD (YEPD): yeast extract, peptone, and dextrose
YPDS: yeast extract, peptone, dextrose, and sorbitol

Pichia pastoris only
BMGY: buffered glycerol-complex medium
BMMY: buffered methanol-complex medium

Minimal Media (also known as drop-out media):
S. cerevisiae
SC (SD): Synthetic complete (YNB, dextrose (or raffinose or galactose), and amino acids)

Pichia pastoris
MGY: minimal glycerol medium
MD: minimal dextrose
MM: minimal methanol
BMGH: buffered minimal glycerol
BMMH: buffered minimal methanol

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Will the Saccharomyces cerevisiae alpha-factor secretion signal be recognized by Schizosaccharomyces pombe?

S. pombe cannot generate P factor when P factor is replaced for alpha in the alpha factor gene. It can, however, produce alpha factor when alpha is replaced for P in the P factor gene. This is negative evidence that S. pombe can process its own mating factor cleavage sites, but not all the cleavage sites of the S. cerevisiae alpha factor. It is better to use a more generic signal sequence (rather than a pre- pro- signal sequence such as alpha). If it is necessary to go the pre- pro- route, it is better to use the S. pombe P factor leader rather than the S. cerevisiae alpha leader.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Do you offer a TOPO-adapted yeast expression vector?

Yes, we do offer the pYES2.1/V5-His-TOPO vector, which is part of the pYES2.1 TOPO TA Expression Kit (Cat. No. K415001), for the direct cloning of Taq polymerase-amplified PCR products and regulated expression in Saccharomyces cerevisiae using galactose.

View all pYES2.1 TOPO™ TA Yeast Expression Kit FAQs

引用および参考文献 (7)

引用および参考文献

Abstract

Identification of a family of animal sphingomyelin synthases.

Authors:Huitema K, Van Den Dikkenberg J, Brouwers JF, Holthuis JC,

Journal:EMBO J

PubMed ID:14685263

'Sphingomyelin (SM) is a major component of animal plasma membranes. Its production involves the transfer of phosphocholine from phosphatidylcholine onto ceramide, yielding diacylglycerol as a side product. This reaction is catalysed by SM synthase, an enzyme whose biological potential can be judged from the roles of diacylglycerol and ceramide as ... More

Cell cycle control of Cdc7p kinase activity through regulation of Dbf4p stability.

Authors:Oshiro G, Owens JC, Shellman Y, Sclafani RA, Li JJ

Journal:Mol Cell Biol

PubMed ID:10373538

'In Saccharomyces cerevisiae, the heteromeric kinase complex Cdc7p-Dbf4p plays a pivotal role at replication origins in triggering the initiation of DNA replication during the S phase. We have assayed the kinase activity of endogenous levels of Cdc7p kinase by using a likely physiological target, Mcm2p, as a substrate. Using this ... More

Regulation of stress response signaling by the N-terminal dishevelled/EGL-10/pleckstrin domain of Sst2, a regulator of G protein signaling in Saccharomyces cerevisiae.

Authors: Burchett Scott A; Flanary Paul; Aston Christopher; Jiang Lixin; Young Kathleen H; Uetz Peter; Fields Stanley; Dohlman Henrik G;

Journal:J Biol Chem

PubMed ID:11940600

'All members of the regulator of G protein signaling (RGS) family contain a conserved core domain that can accelerate G protein GTPase activity. The RGS in yeast, Sst2, can inhibit a G protein signal leading to mating. In addition, some RGS proteins contain an N-terminal domain of unknown function. Here ... More

Genetic screens in yeast to identify mammalian nonreceptor modulators of G-protein signaling.

Authors:Cismowski MJ, Takesono A, Ma C, Lizano JS, Xie X, Fuernkranz H, Lanier SM, Duzic E

Journal:Nature Biotechnology

PubMed ID:10471929

We describe genetic screens in Saccharomyces cerevisiae designed to identify mammalian nonreceptor modulators of G-protein signaling pathways. Strains lacking a pheromone-responsive G-protein coupled receptor and expressing a mammalian-yeast Galpha hybrid protein were made conditional for growth upon either pheromone pathway activation (activator screen) or pheromone pathway inactivation (inhibitor screen). Mammalian ... More

Pheromone-dependent Ubiquitination of the Mitogen-activated Protein Kinase Kinase Ste7.

Authors: Wang Yuqi; Dohlman Henrik G;

Journal:J Biol Chem

PubMed ID:11864977

Many cell signaling pathways are regulated by phosphorylation, ubiquitination, and degradation of constituent proteins. As with phosphorylation, protein ubiquitination can be reversed, through the action of ubiquitin-specific processing proteases (UBPs). Here we have analyzed 15 UBP disruption mutants in the yeast Saccharomyces cerevisiae and identified one (ubp3Delta) that acts specifically ... More

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