本文關(guān)鍵詞： 人博卡病毒(HBoV) HBoV VPU 突變體 sPLA活性　出處：《病毒學(xué)報(bào)》2017年04期 　論文類型：期刊論文

【摘要】：對HBoV2 VP1U蛋白進(jìn)行原核表達(dá)并檢測其sPLA_2活性,對其關(guān)鍵氨基酸進(jìn)行突變,檢測突變對sPLA_2活性的影響。構(gòu)建重組質(zhì)粒pMD18T-VP1U,選取VP1U蛋白4個(gè)關(guān)鍵氨基酸為突變點(diǎn),分別進(jìn)行原核表達(dá),蛋白純化后用sPLA_2試劑盒檢測sPLA_2活性,對比突變前后VP1U蛋白sPLA_2活性有無變化。野生型HBoV2VP1U蛋白的sPLA_2活性為0.243μmol/min/mL,具有sPLA_2活性;突變體L19P、P21R、D42H及Y45N蛋白的sPLA_2活性分別為野生型蛋白的1.2%、1.2%、0.82%、0.41%。野生型HBoV2VP1U蛋白具有sPLA_2活性,突變體蛋白均幾乎完全喪失了sPLA_2活性,提示19、21、42、45位4個(gè)氨基酸是維持sPLA_2活性的關(guān)鍵氨基酸。該研究為靶向抗病毒藥物的研究提供了理論基礎(chǔ)。
[Abstract]:The HBoV2 VP1U protein was expressed in prokaryotic and its sPLA_2 activity was detected, the key amino acids were mutated and the effect of mutation on sPLA_2 activity was detected. The recombinant plasmid pMD18T-VP1U was constructed and four key amino acids of VP1U protein were selected as mutation sites for prokaryotic expression. After purification, sPLA_2 activity was detected by sPLA_2 kit, and the sPLA_2 activity of VP1U protein was compared before and after mutation. The sPLA_2 activity of wild-type HBoV2VP1U protein was 0.243 渭 mol / min / mL, with sPLA_2 activity. The sPLA_2 activity of the mutant L19PmP21RND42H and Y45N protein were 1.2and 1.2and 1.2and 0.82 of wild-type protein, respectively. The wild type HBoV2VP1U protein had sPLA_2 activity, and the mutant protein almost lost sPLA_2 activity. It is suggested that the 4 amino acids at the 45th position of 19s 21 and 42N are the key amino acids to maintain the activity of sPLA_2, which provides a theoretical basis for the study of targeted antiviral drugs.
【作者單位】： 南京醫(yī)科大學(xué)附屬兒童醫(yī)院;
【基金】：江蘇省自然科學(xué)基金青年基金(項(xiàng)目號(hào):BK2012069),題目:HBOV2VP1獨(dú)有區(qū)氨基酸突變對PLA2活性及細(xì)胞因子分泌的影響研究 國家自然科學(xué)基金青年基金(項(xiàng)目號(hào):81300296),題目:HBoV2VP1U突變對PLA2活性及免疫反應(yīng)的影響~~
【分類號(hào)】：R373

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