本文選題：MDV　切入點(diǎn)：UL　出處：《畜牧與獸醫(yī)》2017年05期 　論文類型：期刊論文

【摘要】：選取馬立克氏病毒(MDV)去泛素化酶UL36蛋白N端具有蛋白酶活性的一段氨基酸序列,表達(dá)純化并分析UL36的去泛素化酶活性特點(diǎn)。利用p Cold TF為表達(dá)載體,并在UL36-323的C端連接Strep tagⅡ作為純化標(biāo)簽,構(gòu)建重組質(zhì)粒p Cold TF-UL36-Strep。應(yīng)用大腸桿菌表達(dá)體系,低溫誘導(dǎo)表達(dá),并對(duì)該段蛋白進(jìn)行純化,得到可溶且純度較高的蛋白后,通過(guò)Western blot確定所純化的蛋白即是UL36蛋白。應(yīng)用Di-Ub(K48)和SUMO1-GST作為底物鑒定其去泛素化酶活性。最終得到了可溶且純度較高的UL36-323-Strep蛋白,并且通過(guò)酶切反應(yīng)確定了其去泛素化酶活性,為進(jìn)一步探究UL36在MDV感染及復(fù)制中的作用奠定前期基礎(chǔ),也為探究MDV的致病機(jī)理提供了必要的前提條件。
[Abstract]:A sequence of amino acids with protease activity at the N-terminal of UL36 protein of Marek's virus (MDV) was selected to express, purify and analyze the characteristics of UL36 activity. The expression vector was p Cold TF. The recombinant plasmid p Cold TF-UL36-Strep. expressed in Escherichia coli was induced to express at low temperature, and the protein was purified to obtain soluble and high purity protein, and the recombinant plasmid p#en3# TF-UL36-Strep. was constructed by ligating Strep tag 鈪，

本文編號(hào)：1562460