Grow′n′Glow ACE1 酵母雙雜交系統(tǒng)

品牌：MOBITEC

特點(diǎn)和優(yōu)點(diǎn)：


1. 可以篩選不同蛋白-蛋白之間相互作用的強(qiáng)度；


2. 利用銅誘導(dǎo)的CUP1/ACE1系統(tǒng)能更有效的檢測體內(nèi)蛋白相互作用；


3. 利用銅抗性這種嚴(yán)格的篩選，會(huì)出現(xiàn)很少的假陽性；


4. 利用GFP這種綠色熒光蛋白的報(bào)告基因，篩選陽性菌株可以節(jié)省大量的時(shí)間；


5. 篩選陽性菌株不需要外部酶底物，如X-Gal；


6. 適合高通量的篩選分析。


產(chǎn)品介紹：


酵母雙雜交系統(tǒng)是一種用于篩選體內(nèi)蛋白-蛋白之間相互作用的方法，它依賴于DNA結(jié)合結(jié)構(gòu)域和DNA激活結(jié)構(gòu)域的分離，并將這兩個(gè)結(jié)構(gòu)域基因分別與兩個(gè)蛋白基因融合在一起，如果兩種蛋白能夠相互作用，就會(huì)使這兩個(gè)結(jié)構(gòu)域重組成一個(gè)有活性的轉(zhuǎn)錄因子復(fù)合物，這樣，就會(huì)使報(bào)告基因得以表達(dá)。


MoBiTec的ACE1雙雜交系統(tǒng)是一個(gè)強(qiáng)大的工具，它可以用于新基因的發(fā)現(xiàn)和功能分析，而且可以有效的鑒別體內(nèi)蛋白-蛋白的作用。它將銅誘導(dǎo)的轉(zhuǎn)錄因子ACE1和銅抗性CUP1基因與作為報(bào)告基因綠色熒光蛋白GFP基因結(jié)合起來。兩種相互作用的蛋白分別與ACE1的DNA結(jié)合結(jié)構(gòu)域和DNA激活結(jié)構(gòu)域相融合，從而使我們可以通過下面的方法篩選：1.在含銅的培養(yǎng)基上生長的細(xì)胞；2.依賴銅表達(dá)的強(qiáng)熒光GFP可以通過紫外光檢測到。


因此，這種新技術(shù)，可以很有效的通過一步法篩選可能的陽性表達(dá)，即在含有銅的培養(yǎng)基上生長并發(fā)出綠色熒光的菌株。它是一種快速、高靈敏度和更值得信賴的檢測方法。不斷改變銅濃度還可以篩選出不同蛋白之間相互作用的強(qiáng)度。

hp-Vector表達(dá)系統(tǒng)megaterium芽孢桿菌

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.