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小鼠脂多糖应答蛋白mlrpS的载体构建及融合蛋白表达 被引量:1
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作者 代忠明 陈苏民 +3 位作者 陈南春 杜可军 骆文静 陈景元 《科学技术与工程》 2006年第8期936-938,946,共4页
为构建小鼠mlrpS-cDNA基因原核、真核表达载体,大肠杆菌表达其融合蛋白。采用反转录-聚合酶链反应从经脂多糖刺激的鼠NIH3T3细胞cDNA中,扩增出编码mlrpS的cDNA。用限制性内切酶KpnⅠandXhoⅠ消化后,插入原核表达载体pTAT中,经酶切鉴定... 为构建小鼠mlrpS-cDNA基因原核、真核表达载体,大肠杆菌表达其融合蛋白。采用反转录-聚合酶链反应从经脂多糖刺激的鼠NIH3T3细胞cDNA中,扩增出编码mlrpS的cDNA。用限制性内切酶KpnⅠandXhoⅠ消化后,插入原核表达载体pTAT中,经酶切鉴定与测序证实后,转化大肠杆菌BL21(DE3)菌株。异丙基β-D硫代半乳糖苷(IPTG)诱导产生融合蛋白。经KpnⅠ和XhoⅠ酶切回收mlrpS-cDNA,插入pcDNA3.1载体中;pcDNA3.1-mlrpS再经KpnⅠ和ApaⅠ酶切后,插入到pEGFP-c1载体上,构建pEGFP-mlrpS融合的真核表达载体。构建mlrpS表达载体经测序证实,与GenBank登录的序列完全一致;双酶切鉴定证实,克隆基因正确插入载体pEGFP及pTAT;SDS-PAGE证实融合蛋白表达成功。说明:成功地构建了mlrpS原核、真核表达载体,成功正确表达了6His/mlrpS融合蛋白。 展开更多
关键词 mlrpS 脂多糖应答基因 表达载体 溶核蛋白
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Preparation and Charaterization of Self-assembled Nanoparticles Based on Linolenic-acid Modified Chitosan 被引量:6
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作者 LIU Chenguang Desai Kashappa Goud H. +1 位作者 CHEN Xiguang Park Hyun-Jin 《Journal of Ocean University of China》 SCIE CAS 2005年第3期234-239,共6页
Chitosan was modified by conjugating coupling with linolenic acid through the 1-ethyl-3-(3-dimethylami- nopropyyl) carbodiimide (EDC)-mediated reaction. The degree of substitution 1.8% ( i.e. 1.8 linolenic acid g... Chitosan was modified by conjugating coupling with linolenic acid through the 1-ethyl-3-(3-dimethylami- nopropyyl) carbodiimide (EDC)-mediated reaction. The degree of substitution 1.8% ( i.e. 1.8 linolenic acid group per 100 anhydroglucose units) was measured by ^1H NMR. The critical aggregation concentration (CAC) of the self-aggregate of hydrophobically modified chitosan was determined by measuring the fluorescence intensity of the pyrene as a fluorescent probe. The CAC value in phosphate-buffered saline (PBS) solution (pH 7.4) was 5 × 10^-2 mg mL^-1. The average particle size of selfaggregates of hydrophobically modified chitosan in PBS solution (pH7.4) was 210.8 nm with a unimodal size distribution ranging from 100 to 500 nm. Transmission electron microscopy (TEM) study showed that the formation of near spherical shape nanoparticles has enough structural integrity. The loading ability of hydrophibically modified chitosan (LA-chitosan) was investigated by using bovine serum albumin (BSA) as the model. The loading capacity of self-aggregated nanoparticles increases ( 19.85 % ± 0.04 % to 37.57 % ± 0.25 % ) with the concentration of BSA (0.1-0.5 mg mL^-1 ). 展开更多
关键词 CHITOSAN linolenic acid NANOPARTICLES self-aggregates BSA
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