產(chǎn)品信息

       EZ-Tn5? Transposase* is a hyperactive, muta ted form of Tn5 transposase and a   highly efficient enzyme for insertion of an EZ-Tn5 Trans poson into any target DNA, in vitro.In addition to EZ-Tn5 Transposase, efficient transposition requires that each EZ-Tn5 Transposon have a specific 19-bp transposase recognition se quence (Mosaic End or ME sequence) at each of its ends. Mutations engineered into both the 19-bp MEs and the EZ-Tn5 Transposase results in an in vitro transposition frequency that is 1000-fold greater than wild type.

       EZ-Tn5 Transposase catalyzes a multi-step “cut and paste” transposition reaction. Initially, the enzyme binds the 19-bp ME of   the transposon to form a Transposome?(synaptic complex). The transposome   then randomly attacks and cleaves the phosphodiester backbone of the target   DNA. Finally, the EZ-Tn5 Transposase catalyzes the covalent linkage of the 3′-OH ends of the transposon to the exposed 5′-phos phorylated ends of the target DNA. Transposi tion creates a 9-bp   sequence duplication imme diately flanking the transposon insertion site.

        EZ-Tn5?轉(zhuǎn)座酶是一種高活性，突變形式的Tn5轉(zhuǎn)座酶，是一種在體外將EZ-Tn5轉(zhuǎn)座子插入任何靶標(biāo)DNA的高效酶。除EZ-Tn5轉(zhuǎn)座酶外，高效的轉(zhuǎn)座還要求每個(gè)EZ-Tn5轉(zhuǎn)座子在其每個(gè)末端均具有特定的19 bp轉(zhuǎn)座酶識別序列（Mosaic End或ME序列）。工程化到19 bp的ME和EZ-Tn5轉(zhuǎn)座酶中的突變產(chǎn)生的體外轉(zhuǎn)座頻率是野生型的1000倍。

        EZ-Tn5轉(zhuǎn)座酶催化一個(gè)多步驟的“剪切和粘貼”轉(zhuǎn)座反應(yīng)。 最初，該酶結(jié)合轉(zhuǎn)座子的19 bp ME形成Transposome?（突觸復(fù)合體）。 然后該轉(zhuǎn)座體隨機(jī)攻擊并切割靶DNA的磷酸二酯主鏈。 最后，EZ-Tn5轉(zhuǎn)座酶催化轉(zhuǎn)座子的3'-OH末端與目標(biāo)DNA暴露的5'-磷酸化末端的共價(jià)鍵。 轉(zhuǎn)座可立即形成位于轉(zhuǎn)座子插入位點(diǎn)側(cè)翼的9 bp序列重復(fù)。

        組成成分：

                       EZ-Tn5 Transposase：儲存在-20°C

                       EZ-Tn5 10X Reaction Buffer：儲存在-20°C

                       EZ-Tn5 10X Stop Solution：儲存在-20°C

                       Sterile Water：儲存在-20°C

相關(guān)產(chǎn)品：

TypeOne Restriction Inhibitor（#TY0261H）

TransforMax EC100D pri+ Electrocompetent E.coli（# ECP09500）

TransforMax EC100D pir-116 Electrocompetent E.coli（# EC6P095H） 

參考文獻(xiàn)：

1.   Goryshin, I. Y. and Reznikoff, W. S. (1998) J. Biol. Chem. 273, 7367. 

2.   York, D. et al., (1998) Nucl. Acids Res. 26, 1927.

 技術(shù)參數(shù)

產(chǎn)品應(yīng)用- Insert a marker, T7 promoter, or R6Kγ origin of replication flanked by the 19-bp MEs of Lucigen’s EZ-Tn5 standard Transposons into any target DNA.

- Insert any custom DNA sequence flanked by the 19-bp MEs of an EZ-Tn5 Transposon into any target DNA.

- Prepare Transposomes, in the absence of Mg2+, for electroporation into living bacteria and subsequent random insertion of the transposon into the bacterial chromosome.

- 將Lucigen的EZ-Tn5標(biāo)準(zhǔn)轉(zhuǎn)座子的19 bp ME側(cè)翼的標(biāo)記，T7啟動(dòng)子或R6Kγ復(fù)制起點(diǎn)插入任何目標(biāo)DNA。

- 將EZ-Tn5轉(zhuǎn)座子的19 bp ME側(cè)翼的任何自定義DNA序列插入任何目標(biāo)DNA。

- 在沒有Mg2 +的情況下，準(zhǔn)備轉(zhuǎn)座體以電穿孔進(jìn)入活細(xì)菌，然后將轉(zhuǎn)座子隨機(jī)插入細(xì)菌染色體。