酵母双杂交系统

Grow´n´Glow ACE1 酵母双杂交系统

品牌：MOBITEC

特点和优点：


1. 可以筛选不同蛋白-蛋白之间相互作用的强度；


2. 利用铜诱导的CUP1/ACE1系统能更有效的检测体内蛋白相互作用；


3. 利用铜抗性这种严格的筛选，会出现很少的假阳性；


4. 利用GFP这种绿色荧光蛋白的报告基因，筛选阳性菌株可以节省大量的时间；


5. 筛选阳性菌株不需要外部酶底物，如X-Gal；


6. 适合高通量的筛选分析。


产品介绍：


酵母双杂交系统是一种用于筛选体内蛋白-蛋白之间相互作用的方法，它依赖于DNA结合结构域和DNA激活结构域的分离，并将这两个结构域基因分别与两个蛋白基因融合在一起，如果两种蛋白能够相互作用，就会使这两个结构域重组成一个有活性的转录因子复合物，这样，就会使报告基因得以表达。


MoBiTec的ACE1双杂交系统是一个强大的工具，它可以用于新基因的发现和功能分析，而且可以有效的鉴别体内蛋白-蛋白的作用。它将铜诱导的转录因子ACE1和铜抗性CUP1基因与作为报告基因绿色荧光蛋白GFP基因结合起来。两种相互作用的蛋白分别与ACE1的DNA结合结构域和DNA激活结构域相融合，从而使我们可以通过下面的方法筛选：1.在含铜的培养基上生长的细胞；2.依赖铜表达的强荧光GFP可以通过紫外光检测到。


因此，这种新技术，可以很有效的通过一步法筛选可能的阳性表达，即在含有铜的培养基上生长并发出绿色荧光的菌株。它是一种快速、高灵敏度和更值得信赖的检测方法。不断改变铜浓度还可以筛选出不同蛋白之间相互作用的强度。

hp-Vector表达系统megaterium芽孢杆菌

hp-VectorexpressionSystems for Bacillus megaterium

Features:

High performance vectors with optimized sequence

Protein yield up to 10 times enhanced compared to proteinexpressionwith basic plasmid

Plasmids with established MCS

Encoding C- or N-terminal His-tag for versatile purification (native, 6xHis-tag)

Secretion with LipA or YocH signal peptide up to 9 fold increased

optimized high performance (hp) vectors for Bacillus megaterium!

MoBiTec provides a wide range of useful vectors for the B. megaterium system that are adaptable to most applications and protein purification methods. These include vectors carrying a secretion signal peptide sequence and a 6xHis-tag.


With our new hp-vectors yields up to 10-fold higher compared to the basic plasmids are possible. All plasmids have established multiple cloning sites (MCS) for versatile cloning. Furthermore we offer vectors encoding C- or N-terminal His-tags for easy purification. The protein secretion with LipA or YocH signal peptides is up to nine-fold increased. Induction of proteinexpressionof all vectors is achieved by the tightly regulated and efficiently inducible xylose operon.

Versatile system with a wide range of vectors

The B. megateriumexpressionsystem provides a versatile and easy-to-handle tool for stable and high-yield protein production, both small- and large-scale.


B. megaterium has proven to be an excellent alternative host to E. coli for heterologous geneexpression. Unlike other bacilli strains, proteolytic degradation by alkaline proteases is avoided. In addition, there are no endotoxins found in the cell wall.

B. megaterium protoplasts specifically optimized for transformation

The protoplasts supplied by MoBiTec are from B. megaterium strain WH320, developed by Prof. Dr. W. Hillen at the Institute of Microbiology in Erlangen, Germany. These protoplasts are prepared according to an optimized protocol resulting in the highest transformation efficiencies.


For secretion vectors we offer protoplasts of strain MS941. Both strains are mutants of DSM319, wher WH320 is a chemically mutant, while MS941 has a defined deletion in the gene of the major extracellular protease NprM.








Order Information, shipping and storage

Please click on the respective order # for further details.

order# description amount

BMEG02 Bacillus megaterium protoplasts ready for transformation (strain WH320;1)

Material is sufficient for 4 transformations plus control experiment. 5 x 500 µl

BMEG50 Bacillus megaterium protoplasts, strain MS941 5 x 500 µl

BMEG31 pC-His1623hp shuttle vector, C-term. 6xHis-tag; lyophilized DNA 10 μg

BMEG32 pN-His-TEV1623hp shuttle vector, N-term. His-tag,TEV-site 10 μg

BMEG30 p3STOP1623hp, shuttle vector, lyophilized DNA 10 μg

BMEG33 pSPLipA-hp, shuttle vector with signal sequence, lypophilized DNA 10 μg

BMEG34 pSPYocH-hp, shuttle vector with signal sequence, lypophilized DNA 10 µg


shipped at RT, protoplasts shipped on dry ice

store lyophilized vectors at 4°C, reconstituted vectors at -20°C, protoplasts at -70°C

All vectors are E. coli / B. megaterium shuttle vectors

Please note that protoplasts of Bacillus megaterium have a limited shelf life of some months only, depending on the date of production. Therefore purchase and use of protoplasts must be thoroughly planned.