自動呈色且分子量規則之蛋白質分子量標記套組及其製備方法AUTO-DEVELOPING AND REGULARLY-WEIGHTED PROTEIN MOLECULAR WEIGHT MARKER KIT AND METHOD FOR PREPARING THE SAME

專利類型

發明

專利國別 (專利申請國家)

美國

專利申請案號

13/402,086

專利證號

US 8,791,240 B2

專利獲證名稱

自動呈色且分子量規則之蛋白質分子量標記套組及其製備方法AUTO-DEVELOPING AND REGULARLY-WEIGHTED PROTEIN MOLECULAR WEIGHT MARKER KIT AND METHOD FOR PREPARING THE SAME

專利所屬機關 (申請機關)

高雄醫學大學

獲證日期

2014/07/29

技術說明

我們開發的14-tags自動呈色且分子量規則的蛋白分子量標記是將目前於蛋白質研究上所廣泛使用的14個tag-markers，再依序接上maltose-binding protein (MBP)、Glutathione-S-transferase(GST)、 thioredxin (TRX)，以形成分子量規則之重組蛋白標記，因為每個標記均包含有14-Tags，所以，當研究者使用Anti-Tags antibody來偵測他們的蛋白質時，14-tags蛋白標記即會自動呈現於底片上。我們已表現並純化不同分子量的14-tags蛋白分子量標記，藉由comassie blue染色，發現此14-tags蛋白分子量標記確實能規則性呈現於SDS-PAGE上，且分子量準確；而西方點墨法亦證實，此14-tags蛋白分子量標記確實能規則性顯影於底片上。我們研發的14-tags 蛋白分子量標記具有以下優點: :(1)方便性:可以伴隨著1級或2級抗體染色自動將marker呈現在底片上，不需要再手劃marker位置，且此14-tags 蛋白分子量標記亦可當作Anti-Tags Ab的陽性控制組相當實用。(2)準確性:此蛋白分子量標記是dye-free，所以分子量準確。(3)可呈現分子量規則的Ladder band，易於觀測、判斷。 We proposes to develop an innovative protein molecular weight marker which incorporate 14 commonly used tags (14-tag). This innovative protein molecular weight marker will offer the benefit of auto-developing and regular-weighted. In our preliminary results, we show that the “14-tag” recombinant protein can be expressed in the BL21 competent cells and subsequently purified by nickel columns. Importantly, the purified “14-tag” recombinant protein can be recognized by different anti-tag (HA, myc, Flag, V5 etc.) antibodies. Based in these results, we propose to use the “14-tag” as basic recognition units, and fuse them to TRX, GST and MBP to regularly increase the molecular weights of the recombinant protein marker. Owing to the intramolecular tags, the proposed protein molecular weight marker will provide the convenience of auto-developing on the film when the investigators use anti-tags antibodies in the detection of their target proteins in the western blot.

備註

連絡單位 (專責單位/部門名稱)

智財保護與科技管理組

連絡電話

07-3138030

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