試劑中英文名稱:Zeocin (100mg/ml) 博萊霉su(100 mg/ml)
儲(chǔ)存:-20℃干燥避光保存至少1年,冰袋運(yùn)輸。
化學(xué)式:C55H83N19O21S2Cu
Zeocin is a formulation of phleomycin D1, a basic, water-soluble, copperchelated glycopeptide isolated from Streptomyces verticillus and shows strong toxicity against bacteria, fungi (including yeast), plants, and mammalian cell lines.
The blue color of the solution is due to the presence of copper and the copper-chelated form of Zeocin is inactive.
When the antibiotic enters the cell, the copper cation is reduced from Cu2+ to Cu+ and removed by sulfhydryl compounds in the cell.
Upon copper removal, Zeocin is activated, and binds and cleaves DNA, causing cell death.
A Zeocin resistance protein of 13,665 Da, has been isolated and characterized. The protein is the product of the Sh ble gene, binds stoichiometrically to Zeocin and inhibits its DNA strand cleavage activity.
Expression of this protein in eukaryotic and prokaryotic hosts confers resistance to Zeocin.
Zeocin 是葉霉素 D1 的制劑,葉霉素 D1 是一種堿性、 水溶性、 銅螯合糖肽,從對(duì)細(xì)菌、 真菌 (包括酵母 )、 植物和哺乳動(dòng)物細(xì)胞具有較強(qiáng)的毒性。
溶液的藍(lán)色是由于銅的存在,并且 Zeocin 的銅螯合形式?jīng)]有活性。
當(dāng)抗生素進(jìn)入細(xì)胞時(shí),銅陽(yáng)離子從 Cu2+ 還原為 Cu+,并被巰基化合物去除。
銅被去除后,Zeocin 被激活,結(jié)合并切割 DNA,導(dǎo)致細(xì)胞死亡。
Zeocin 耐藥性13,665Da 的蛋白質(zhì),已被分離和表征。
該蛋白質(zhì)是 Shble 基因的產(chǎn)物,與 Zeocin 化學(xué)計(jì)量結(jié)合并抑制其 DNA 鏈切割活動(dòng)。
這種蛋白質(zhì)在真核和原核宿主中的表達(dá)賦予了對(duì) Zeocin 的抗性。
穩(wěn)定轉(zhuǎn)染細(xì)胞系的維持培養(yǎng)
可采取以下方式維持培養(yǎng)穩(wěn)定轉(zhuǎn)染細(xì)胞:
1)使用含相同劑量Zeocin的篩選培養(yǎng)基來維持培養(yǎng);
2)降低Zeocin劑量為原來的一半進(jìn)行維持培養(yǎng);
3)使用正好能預(yù)防敏感細(xì)胞生長(zhǎng)但不足以致死的Zeocin劑量來維持培養(yǎng)【根據(jù)殺滅曲線來判斷】。
二、使用優(yōu)勢(shì)(Zeocin博萊霉su)
100mg/ml即用型無菌溶液,可直接加入培養(yǎng)基,省去配制步驟。
提供凍干粉(需HEPES緩沖液重溶)及預(yù)溶母液兩種形式,適應(yīng)不同需求。
單一抗生素適用于原核(細(xì)菌)、真核(酵母、哺乳動(dòng)物細(xì)胞)篩選,簡(jiǎn)化實(shí)驗(yàn)流程。
抗性細(xì)胞形態(tài)與正常細(xì)胞無異,便于表型觀察。
通過濃度梯度實(shí)現(xiàn)選擇性殺傷,Sh ble抗性基因提供明確篩選標(biāo)記。
維持培養(yǎng)時(shí)可靈活調(diào)整濃度,平衡細(xì)胞生長(zhǎng)與抗性維持。
用于科研相關(guān)領(lǐng)域,不能用于人體實(shí)驗(yàn)!
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