蛋白質(zhì)酪氨酸磷酸酶MEG1催化結(jié)構(gòu)域的克隆表達(dá)及其多克隆抗體的制備
發(fā)布時(shí)間:2019-06-24 13:30
【摘要】:為了研究PTPMEG1的催化結(jié)構(gòu)域在細(xì)胞信號(hào)轉(zhuǎn)導(dǎo)中的功能,我們以pBluescriptⅡKS PTPMEG1質(zhì)粒為模板,首先克隆了PTPMEG1催化結(jié)構(gòu)域基因ΔPTPMEG1,構(gòu)建了pT7-ΔPTPMEG1質(zhì)粒。通過轉(zhuǎn)化克隆宿主大腸桿菌DH5α進(jìn)行擴(kuò)增后,提取質(zhì)粒,轉(zhuǎn)化表達(dá)宿主大腸桿菌RosetaDE3,通過IPTG誘導(dǎo),進(jìn)行ΔPTPMEG1的原核表達(dá),確定該表達(dá)為包涵體表達(dá),利用制備電泳分離純化,獲得了純化后的ΔPTPMEG1蛋白。用純化后的蛋白對(duì)家兔進(jìn)行免疫,制各ΔPTPMEG1的多克隆抗體,并利用“PVDF固定抗原親和層析法”對(duì)ΔPTPMEG1多克隆抗體進(jìn)行純化,得到了純度、效價(jià)及靈敏度較高的ΔPTPMEG1抗體,為以后對(duì)PTPMEG1在細(xì)胞中的生理功能和組織分布的研究奠定了基礎(chǔ)。
[Abstract]:In order to study the function of PTPMEG1 catalytic domain in cell signal transduction, we first cloned PTPMEG1 catalytic domain gene 螖 PTPMEG1, and constructed pT7- 螖 PTPMEG1 plasmid using pBluescript 鈪,
本文編號(hào):2505092
[Abstract]:In order to study the function of PTPMEG1 catalytic domain in cell signal transduction, we first cloned PTPMEG1 catalytic domain gene 螖 PTPMEG1, and constructed pT7- 螖 PTPMEG1 plasmid using pBluescript 鈪,
本文編號(hào):2505092
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