盐穗木HcUKPP原核表达及重组菌非生物胁迫耐受性研究
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引用本文:张丽丽,张 冀,张富春.盐穗木HcUKPP原核表达及重组菌非生物胁迫耐受性研究[J].西北植物学报,2018,38(2):219~224
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作者单位
张丽丽,张 冀,张富春* (新疆大学 生命科学与技术学院新疆生物资源基因工程重点实验室乌鲁木齐 830046) 
基金项目:新疆重点实验室专项资金(2014KL001)
中文摘要:该研究采用RT PCR技术,从盐穗木cDNA文库中克隆获得未知功能多肽HcUKPP基因,构建了大肠杆菌Escherichia coli BL21∷pET30a HcUKPP重组菌株,并检测了重组菌株在不同非生物胁迫下的耐受性。结果显示:HcUKPP基因开放阅读框为243 bp,融合His的HcUKPP蛋白的分子量约为15 kD。在37 ℃条件下,不同浓度的异丙基 β D硫代半乳糖苷(IPTG)诱导4 h后His HcUKPP融合蛋白均可表达,且E.coli BL21∷pET30a HcUKPP重组菌在不同浓度NaCl(100~900 mmol/L)、聚乙二醇(2.5%~20%,PEG 6000)和甲基紫精(25~200 μmol/L)胁迫处理下,其生长均具有明显优势。尤其是在500 mmol/L NaCl、10% PEG 6000和75 μmol/L甲基紫精胁迫12 h后,重组大肠杆菌BL21呈现出极显著的优势,分别达到了对照菌的1.81、1.47和3.48倍。研究表明,盐穗木HcUKPP可以显著提高重组大肠杆菌对不同非生物胁迫的耐受性,证明HcUKPP是一类新发现的能够响应非生物胁迫的多肽。
中文关键词:盐穗木  HcUKPP  原核表达  重组大肠杆菌  非生物耐受
 
Procaryotic Expression of HcUKPP from Halostachys caspica and Analysis of Abiotic Stress Tolerance of the Recombinant Bacteria
Abstract:The recombinant strain Escherichia coli BL21∷pET30a HcUKPP was obtained by the method of prokaryotic vector construction after cloning of unknown functional peptide HcUKPP gene, and then tested its tolerance under different abiotic stresses. The results revealed that open reading frame(ORF) of the gene was 243 bp, the molecular weight of the recombinant HcUKPP was approximately 15 kD. In addition, the expression of His HcUKPP fusion protein could be induced with different concentrations of isopropyl β D 1 thiogalactopyranoside (IPTG) for 4 h at 37 ℃. Furthermore, the growth of recombinant strain E.coli BL21∷pET30a HcUKPP have shown obvious advantages under treatment of different concentrations of NaCl (100-900 mmol/L), polyethylene glycol (2.5%-20%, PEG 6000) and methyl viologen (25-200 μmol/L). Especially under the condition of 500 mmol/L NaCl, 10% PEG 6000 and 75 μmol/L methyl viologen for 12 h, interestingly,recombinant E.coli BL21 showed significant advantages, their growth reached 1.81, 1.47 and 3.48 fold of control bacteria respectively. Overall, the HcUKPP gene of Halostachys caspica can significantly improve tolerance of recombinant E.coli BL21 under different abiotic stresses, which proved that the HcUKPP is a kind of newly discovered polypeptide in response to abiotic stress.
keywords:Halostachys caspica  unknown functional polypeptide  prokaryotic expression  recombinant Escherichia coli  abiotic tolerance
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