Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM.
Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM.
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嗜酸乳杆菌NCFM沉默slpB的分子克隆、表达及粘附分析
DOI：
10.1186/s13568-018-0631-2
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发表时间：
2018-06-23
期刊：
影响因子：
3.7
通讯作者：
Pan D
中科院分区：
文献类型：
--
作者：
Guo Y;Li X;Yang Y;Wu Z;Zeng X;Nadari F;Pan D
成功地扩增了嗜酸乳杆菌NCFM的slpB基因，并将其连接到重组载体pET-28 a的相应位点上。然后将pET-28 a-slpB载体转化到大肠杆菌DH（DE 3）中，并通过在37 °C下用ImM IPTG诱导14小时来表达融合His-slpB蛋白。所得His-slpB蛋白（SB）具有48 kDa的相对分子量。经Ni-NTA柱纯化，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和western blot对比分析证实。从L.嗜酸乳杆菌NCFM的提取和纯化。其相对分子量为46 kDa。圆二色性分析表明，两种S层蛋白的β-折叠含量高，α-螺旋含量低。在粘附实验中，SA对Caco-2细胞的粘附能力高于SB。结果表明，这两个S层蛋白可能具有生物技术应用。
The slpB gene of Lactobacillus acidophilus NCFM, which differs from the slpA gene and is silent under normal conditions, was successfully amplified and ligated to the corresponding available sites on a recombinant pET-28a vector. Then the pET-28a-slpB vector was transformed into Escherichia coli DH (DE3) and the fusion His-slpB protein was expressed by induction with 1 mM IPTG for 14 h at 37 °C. The resulting His-slpB protein (SB) had a relative molecular weight of 48 kDa. It was purified using a Ni-NTA column and was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot contrastive analysis. The slpA protein (SA) from L. acidophilus NCFM was extracted and purified. It had a relative molecular weight of 46 kDa. Circular dichroism measurements suggested that the two S-layer proteins had a high β-sheet content and a low α-helix structure content. In an adhesion experiment, SA displayed higher adhesive capability towards Caco-2 cells than did SB. The results suggest that these two S-layer proteins could have biotechnological applications.
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影响因子：
4.4
作者：
Lindholm, A;Smeds, A;Palva, A
通讯作者：
Palva, A
影响因子：
11.4
作者：
Lightfoot, Yaima L.;Selle, Kurt;Mohamadzadeh, Mansour
通讯作者：
Mohamadzadeh, Mansour
DOI：
10.1186/1476-9255-9-7
发表时间：
2012-03-16
期刊：
Journal of inflammation (London, England)
影响因子：
--
作者：
Zadeh M;Khan MW;Goh YJ;Selle K;Owen JL;Klaenhammer T;Mohamadzadeh M
通讯作者：
Mohamadzadeh M
影响因子：
3.2
作者：
Hynönen, U;Westerlund-Wikström, B;Korhonen, TK
通讯作者：
Korhonen, TK
影响因子：
2.1
作者：
Novotny, R;Scheberl, A;Schäffer, C
通讯作者：
Schäffer, C