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冻干人用狂犬病疫苗(Vero细胞)蛋白结构特性分析
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作者 匡志新 柴博雅 +7 位作者 李春艳 孙宏亮 朱美宣 熊晋峰 谢冰丽 王帆 解辉 邹勇 《中国生物制品学杂志》 CAS CSCD 2019年第4期375-379,共5页
目的分析冻干人用狂犬病疫苗(Vero细胞)蛋白的结构特性。方法 SDS-PAGE电泳分离4批冻干人用狂犬病疫苗(Vero细胞)原液,电泳条带经nano LC-MS/MS进行蛋白鉴定,Native-PAGE分析病毒颗粒的完整性,并分析蛋白相对分子质量。同时进行N-末端... 目的分析冻干人用狂犬病疫苗(Vero细胞)蛋白的结构特性。方法 SDS-PAGE电泳分离4批冻干人用狂犬病疫苗(Vero细胞)原液,电泳条带经nano LC-MS/MS进行蛋白鉴定,Native-PAGE分析病毒颗粒的完整性,并分析蛋白相对分子质量。同时进行N-末端氨基酸序列分析。结果 SDS-PAGE分析可见5个主条带,分别为狂犬病毒G、M、N、P蛋白及G蛋白二聚体(G-1),经nano LC-MS/MS鉴定,确定相对分子质量分别为74 000、21 885、57 287、40 934及141 067,各结构蛋白均包含在病毒颗粒中,M、N、P、G蛋白的N-末端起始氨基酸序列为分别为MNFLR、MDADR、MSKIF、MVPQA,与预期一致。结论 4批冻干人用狂犬病疫苗(Vero细胞)原液所含结构蛋白源于狂犬病病毒。 展开更多
关键词 狂犬病疫苗 结构蛋白 SDS-PAGE分析 nano LC-MS/MS 氨基酸序列分析 VERO细胞
猪链球菌Ide通用截短蛋白原核表达及纯化研究 预览
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作者 曲光刚 王长江 +5 位作者 李书光 李茂峰 武曰星 金婷婷 韩文瑜 沈志强 《中国兽药杂志》 2018年第6期7-12,共6页
为利用大肠埃希菌表达猪链球菌Ide通用截短蛋白,通过对GenBank公布的IdeS蛋白家族基因同源性分析,使用Primer 5.0软件设计一对特异性引物,以猪链球菌2型强毒株JZLQ全基因组为模板,采用PCR方法扩增Ide基因截短片段,经 Bam HⅠ和 Hi... 为利用大肠埃希菌表达猪链球菌Ide通用截短蛋白,通过对GenBank公布的IdeS蛋白家族基因同源性分析,使用Primer 5.0软件设计一对特异性引物,以猪链球菌2型强毒株JZLQ全基因组为模板,采用PCR方法扩增Ide基因截短片段,经 Bam HⅠ和 Hin dⅢ双酶切后将目的片段分别克隆到pET28a、pET32a和pET-sumo表达载体上,分别转化宿主菌 E.coli BL21(DE3)。通过优化诱导温度及诱导剂IPTG浓度进行表达并对表达产物进行SDS-PAGE分析。结果显示,PCR扩增片段大小约为1200 bp,经双酶切和测序验证构建正确;三种重组表达载体转化大肠埃希菌后均有目的蛋白表达,但不同表达条件下目的蛋白表达量存在差异,其中pET28a-TrIde重组表达载体用25 ℃诱导、IPTG终浓度为1 mmol/L时可实现重组蛋白的高效可溶性表达,为Ide Ssuis 蛋白在猪链球菌通用疫苗研发方面的研究奠定了基础。 展开更多
关键词 猪链球菌2型 IgM裂解酶 原核表达 SDS-PAGE分析
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糙皮侧耳脲酶基因的克隆和原核表达分析
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作者 文晴 邹明 +3 位作者 靳橄 郭丹丹 魏忠方 申进文 《菌物学报》 CSCD 北大核心 2018年第11期1498-1506,共9页
尿素是现代农业生产中应用较为广泛的一种氮素化肥,而脲酶(EC3.5.1.5)是尿素分解利用的关键酶。本文以糙皮侧耳Pleurotus ostreatus栽培菌株New 831为试验材料,探究了糙皮侧耳对尿素的利用情况,结果表明:糙皮侧耳可利用尿素作为唯一... 尿素是现代农业生产中应用较为广泛的一种氮素化肥,而脲酶(EC3.5.1.5)是尿素分解利用的关键酶。本文以糙皮侧耳Pleurotus ostreatus栽培菌株New 831为试验材料,探究了糙皮侧耳对尿素的利用情况,结果表明:糙皮侧耳可利用尿素作为唯一氮源,平板培养时尿素最适添加量为20mmol/L;在液体摇瓶培养过程中,培养液中的铵根浓度表现为先急剧升高后缓慢降低。通过对P. ostreatus PC15菌株基因组分析,获得了一个功能注释为脲酶的基因,并克隆获得了其全长基因组DNA(gDNA)和编码区(CDS)片段,命名为Pourease。结果表明:Pourease基因的gDNA和CDS长度分别为3 003bp和2 517bp,由10个外显子和9个内含子组成;Pourease蛋白由838个氨基酸组成,预测分子量为90.03kDa,与SDS-PAGE分析结果相符;Pourease蛋白与细菌、真菌和植物来源的脲酶具有52%–82%的一致性,且含有脲酶保守的镍离子结合位点;与洋刀豆脲酶空间结构类似,Pourease蛋白也以同源三聚体的形式存在。 展开更多
关键词 糙皮侧耳 尿素 铵根 脲酶 SDS-PAGE分析 同源三聚体
太子参蛋白提取工艺优化及SDS-PAGE分析 预览
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作者 晋海军 王海霞 +1 位作者 张涛 向守艳 《四川农业大学学报》 CSCD 北大核心 2018年第4期500-506,共7页
【目的】研究太子参的最佳蛋白提取工艺和不同参形太子参蛋白条带差异。【方法】选取3种蛋白提取方法(Tris-HCl法、TCA-丙酮法、饱和酚法)对贵州施秉县牛大场太子参种植基地的3种参形太子参进行蛋白提取及SDSPAGE分析,利用响应面分析法... 【目的】研究太子参的最佳蛋白提取工艺和不同参形太子参蛋白条带差异。【方法】选取3种蛋白提取方法(Tris-HCl法、TCA-丙酮法、饱和酚法)对贵州施秉县牛大场太子参种植基地的3种参形太子参进行蛋白提取及SDSPAGE分析,利用响应面分析法对筛选出的最佳蛋白提取方法的提取工艺进行优化。【结果】3种蛋白提取方法中Tris-HCl法提取的蛋白含量较高,经SDS-PAGE分析,太子参蛋白条带大致分布在10kDa~90kDa之间,15kDa处一等参的蛋白表达量明显高于二等参和统货。利用响应面优化法获得提取太子参蛋白的最佳工艺。【结论】获得太子参蛋白的最佳提取工艺:料液比1:1.32、提取温度2 5℃、pH值7.47、提取时间1.5 h,还发现不同参形太子参蛋白在15kDa处的表达量存在明显差异,这可为今后利用蛋白质组学技术进一步揭示不同参形太子参形成的分子机制提供理论依据。 展开更多
关键词 太子参 参形 SDS-PAGE分析 响应面优化
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用电气泳动分析小麦粉中的蛋白质
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作者 宋军 周远 《粮食加工》 2017年第6期13-14,共2页
对市场上不同厂家的多种小麦粉中的水分、灰分、蛋白质、谷蛋白湿重量和谷蛋白的干含量、粉色、粉粒大小分布、蛋白质分子大小的分布进行了分析。小麦样品抽提物的SDS-PAGE分析结果:产品间的差异表现在两种分子量为30 k D麦醇溶蛋白的... 对市场上不同厂家的多种小麦粉中的水分、灰分、蛋白质、谷蛋白湿重量和谷蛋白的干含量、粉色、粉粒大小分布、蛋白质分子大小的分布进行了分析。小麦样品抽提物的SDS-PAGE分析结果:产品间的差异表现在两种分子量为30 k D麦醇溶蛋白的上面。产品间存在差异的麦醇溶蛋白存在于小麦外皮的附近,它可以用做一种小麦制粉指标。 展开更多
关键词 小麦粉中 蛋白质 SDS-PAGE分析 麦醇溶蛋白
Inducible Expression on Multi-epitope of Porcine Circovirus Type 2 and Its Immunological Competence 预览
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作者 Dong Lin Wang Yanping +3 位作者 Wang Jinliang Mo Ling Shen Zhiqiang Liu Zengshan 《动物与饲料科学:英文版》 CAS 2016年第3期151-154,158共5页
In order to obtain induction expressed porcine circovirus type 2(PCV2)multi-epitope with good immunogenicity in vitro.Major epitopes of PCV2 were screened in the test,epitopes were composed sequentially,the c DNA sequ... In order to obtain induction expressed porcine circovirus type 2(PCV2)multi-epitope with good immunogenicity in vitro.Major epitopes of PCV2 were screened in the test,epitopes were composed sequentially,the c DNA sequences were artificially synthesized.Bam HⅠand SalⅠwere directionally cloned into prokaryotic expression vector PEGX-4T-1 multiple cloning site,then BL21 competent cells were transformed,positive clones were screened,IPTG inducible expression was conducted.Expression on target gene was analyzed by SDS-PAGE,fusion protein polypeptide was extracted and purified,immunocompetence of the expressed multi-epitope protein was identified by Westernblot.BALB/c mouse was immuned by fusion protein polypeptide,the antibody was determined by ELISA,immunogenicity was evaluated.Results showed that expression recombinant plasmid pEGX-4T-1-ep contained with seven PCV2 antigen epitopes had been constructed successfully.SDS-PAGE analysis showed that fusion protein polypeptide was expressed effectively in Escherichia coli(E.coli),and the molecular weight was about 35ku,which existed in the form of solubility.Results of Westernblot showed that the extraction and purification of fusion protein polypeptide and PCV2 positive serum had good reactogenicity.Results of ELISA showed that the purified fusion protein polypeptide could stimulate the body to produce PCV2 specific antibody which had good immunogenicity.Results indicated that the constructed PCV2 multi-epitope had good expression characteristics in vitro,and the expression protein had good immunogenicity.The study provided a basic for the study on PCV2 epitope screening,functional identification and multi-epitope vaccine. 展开更多
关键词 猪圆环病毒2型 诱导表达 免疫能力 多表位 SDS-PAGE分析 SDS-PAGE 蛋白多肽 PCV2
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从海洋酵母羧甲基纤维素酶(出芽短梗霉98):纯化,鉴定,基因克隆和羧甲基纤维素的消化 预览
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作者 RONG Yanjun ZHANG Liang +1 位作者 CHI Zhenming WANG Xianghong 《中国海洋大学学报:英文版》 SCIE CAS 2015年第5期913-921,共9页
我们曾报道了短梗霉菌产生的高纤维素酶产量98。在这项研究中,羧甲基纤维素酶(CMCase)在培养的细胞。短梗霉98的纯化至均一,与酶的最大产量为4.51 U(mg蛋白)- 1。SDS-PAGE分析表明,纯化的酶的分子量为67.0kda。具有相当的敏感性... 我们曾报道了短梗霉菌产生的高纤维素酶产量98。在这项研究中,羧甲基纤维素酶(CMCase)在培养的细胞。短梗霉98的纯化至均一,与酶的最大产量为4.51 U(mg蛋白)- 1。SDS-PAGE分析表明,纯化的酶的分子量为67.0kda。具有相当的敏感性为40℃纯化的酶的最适温度,比从其他真菌的cmcases低得多。该酶的最佳pH值为5.6,和活动的个人资料被稳定在一个范围内的酸度(pH 5,0-6.0)。这种酶被激活Na+,Mg2+,Ca2+,K+,Fe2+和Cu2+,然而,它是由Fe3+,Ba2+,Zn2+,Mn2+和银离子抑制。公里和纯化的酶的Vmax值4.7mgml-1 0.57 pmol L-1 min-1(mg蛋白)1,分别。只有大小不同的低聚糖,羧甲基纤维素(CMC)释放与纯化的酶水解后。该基因编码的酶是A.霉98个克隆,其中包含一个开放阅读框(eu978473)意义。推导的蛋白质含有酶超家族的保守结构域(糖基水解酶家族5)。的N-末端氨基酸序列的纯化的酶是m-a-p-h-a-e-p-q-s-q-t-t-e-q-t-s-s-g-q-f,这与从克隆的基因推导一致。这表明,纯化的酶是由克隆纤维素酶基因在酵母编码。 展开更多
关键词 羧甲基纤维素酶 出芽短梗霉 基因克隆 海洋酵母 纯化 SDS-PAGE分析 最佳pH值 鉴定
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Gene Cloning and Molecular Characterization of a β-Glucosidase from Thermotoga Naphthophila RUK-10:an Effective Tool for Synthesis of Galacto-oligosaccharide and Alkyl Galactopyranosides
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作者 KONG Fansi YANG Jingwen +4 位作者 ZHEN Zhen LIANG Tingting ZHU Dongliang GAO Renjun XIE Guiqiu 《高等学校化学研究:英文版》 SCIE CAS CSCD 2015年第5期774-780,共7页
关键词 β-葡萄糖苷酶 半乳糖寡糖 烷基糖苷 基因克隆 分子特性 合成 SDS-PAGE分析 水解活性
Co-expression of Clostridium perfringens α and ε Toxins and Their Immunogenicity 预览
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作者 Han Guangwei Li Zhaocai +5 位作者 Gong Xiaowei Chen Qiwei Cao Xiaoan Zheng Fuying Zhang Xiaoying Zhou Jizhang 《动物与饲料科学:英文版》 CAS 2015年第2期74-77,共4页
The gene CPA and ETX encoding α and ε toxins were amplified from the standard strain C60-2 of type D Clostridium perfingens by PCR,and then they were inserted into pet-Duet-1 vecto,respectively for the construction ... The gene CPA and ETX encoding α and ε toxins were amplified from the standard strain C60-2 of type D Clostridium perfingens by PCR,and then they were inserted into pet-Duet-1 vecto,respectively for the construction of co-expression plasmid p ETDuet-1-CPA-ETX. The co-expression plasmid was transformed into BL21( DE3) competent cells. α and ε toxins proteins were induced by IPTG before analysis by SDS-PAGE. As a result,both of α and ε toxins were detected at 43 and 34 KU by western-blot. Mice immunized with the co-expressed α and ε toxins proteins produced high titers of neutralizing antibodies in the serum,which protected the mice against the attack of type D C. perfringens culture filtrate. In addition,mice immunized with the produced co-expressed α and ε toxins proteins showed significantly higher surviving rate than with ε toxins protein alone when infected with culture filtrate of type D C. perfringens. These results indicated that co-expression of α and ε toxins proteins could be used as a new method to prepare vaccines against the pathogens of multiple serotypes. 展开更多
关键词 产气荚膜梭菌 共表达质粒 毒素蛋白 免疫原性 SDS-PAGE分析 免疫小鼠 培养滤液 PCR扩增
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Characterization of a Bacteriocin-Like Substance Produced from a Novel Isolated Strain of Bacillus subtilis SLYY-3 预览
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作者 LI Junfeng LI Hongfang +2 位作者 ZHANG Yuanyuan DUAN Xiaohui LIU Jie 《中国海洋大学学报:英文版》 SCIE CAS 2014年第6期995-999,共5页
In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY-3, which produced a bacteriocin-like substance(BLS), was characterized to be a strain of Bacillus... In the present research, the strain SLYY-3 was isolated from sediments of Jiaozhou Bay, Qingdao, China. The strain SLYY-3, which produced a bacteriocin-like substance(BLS), was characterized to be a strain of Bacillus subtillis by biochemical profiling and 16 S r DNA sequence analysis. It is the first time to report that Bacillus subtilis from Jiaozhou Bay sediments could produce a BLS. The BLS of B. subtillis SLYY-3 exhibited strong inhibitory activity against gram-positive bacteria(including Staphylococcus aureus and B. subtillis) and some fungi(including Penicillium glaucum, Aspergillus niger and Aspergillus flavus). The antimicrobial activity was detected from culture in the exponential growth phase and reached its maximum when culture entered into stationary growth phase. It was thermo-tolerant even when being kept at 100℃ for 60 min without losing any activity and stable over a wide p H range from 1.0 to 12.0 while being inactivated by proteolytic enzyme and trypsin, indicating the proteinaceous nature of the BLS. The BLS was purified by precipitation with hydrochloric acid(HCl) and gel filteration(Sephadex G-100). SDS-PAGE analysis of the extracellular peptides of SLYY-3 revealed a bacteriocin-like protein with a molecular mass of 66 k Da. Altogether, these characteristics indicate the potential of the BLS for food industry as a protection against pathogenic and spoilage microorganisms. 展开更多
关键词 枯草芽孢杆菌 细菌素 分离株 物质 SDS-PAGE分析 RDNA序列分析 金黄色葡萄球菌 葡聚糖凝胶
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Construction of a Food-Grade Expression Vector Based on pMG36e by Using an α-Galactosidase Gene as a Selectable Marker 预览
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作者 GU Xin-xi TAN Jian-xin +3 位作者 TIAN Hong-tao ZHANG Yu-lan LUO Yun-bo GUO Xing-hua 《农业科学学报:英文版》 SCIE CSCD 2014年第8期1802-1808,共7页
Construction of a food-grade expression vector for application to lactic acid bacteria(LAB) is of importance for dairy fermentation system. An α-galactosidase(aga) gene encoding an enzyme degrading melibiose was ampl... Construction of a food-grade expression vector for application to lactic acid bacteria(LAB) is of importance for dairy fermentation system. An α-galactosidase(aga) gene encoding an enzyme degrading melibiose was amplified by PCR from the plasmid p RAF800 of Lactococcus lactis NZ9000. The aga gene was introduced into pMG36 e to substitute the p rimary antibiotic selectable marker of pMG36 e, resulting in construction of a new food-grade expression vector pMG36-aga. To testify the expression efficiency of exogenous gene in pMG36-aga, a 1.5 kb long α-amylase(amy) gene from Ba cillus li cheniformis was cloned by PCR and introduced into the plasmid pMG36-aga. The resultant plasimd pMG36-aga-amy was transformed into L. lactis ML23 by electroporation. The positive clones were selected with the medium containing melibiose as the sole carbon source. Th e selection efficiency of aga was 8.71×103 CFU with a standard deviation of 9.1×102 CFU ?g-1 DNA of pMG36-aga. Furthermore, the SDS-PAGE analysis showed that the pMG36-aga-amy expressed a 56.4 kDa protein which was the same as the putati ve molecular weight of α-amylase. The starch plate assay also indicated that L. lactis ML23 displayed high activity of α-amylase by expressing of amy gene of pMG36-aga-amy. 展开更多
关键词 半乳糖苷酶基因 表达载体 选择标记 食品级 SDS-PAGE分析 乳酸乳球菌 PCR扩增 PCR克隆
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畜牧兽医科技文摘 预览
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《中国畜牧兽医文摘》 2013年第5期201-210,共10页
将猪瘟病毒不同抗原表位基因串联构成重组基因BT21,化学合成后克隆至pMD18-T载体中,然后再将BT21串联基因片段插入原核表达载体pGEX-6P-1。经酶切和测序鉴定后,构建的重组质粒pGEX-BT21转化大肠杆菌BL21(DE3),通过IPTG诱导表达,... 将猪瘟病毒不同抗原表位基因串联构成重组基因BT21,化学合成后克隆至pMD18-T载体中,然后再将BT21串联基因片段插入原核表达载体pGEX-6P-1。经酶切和测序鉴定后,构建的重组质粒pGEX-BT21转化大肠杆菌BL21(DE3),通过IPTG诱导表达,表达产物进行SDS-PAGE分析,用GlutathioneSepharose4B亲和层析法纯化目的蛋白和Westernblot分析其免疫学活性。结果表明:融合蛋白GST-BT21以可溶形式表达,分子量约为33kDa,与预期大小相符,纯化的重组蛋白可被猪瘟病毒阳性血清所识别,具有良好的免疫学活性。从而为进一步研究该融合蛋白的免疫特性和功能奠定了基础。 展开更多
关键词 兽医科技 SDS-PAGE分析 免疫学活性 文摘 畜牧 原核表达载体 BLOT分析 基因串联
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Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column
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作者 SHEN Shaochuan WANG Liangyan +5 位作者 SUN Zongtao LI Mingfeng LIU Chengzhi TIAN Bing YUN Junxian HUA Yuejin 《中国化学工程学报:英文版》 SCIE EI CAS CSCD 2013年第6期663-669,共7页
关键词 耐辐射球菌 细胞工程 层析分离 二磷酸 SDS-PAGE分析 亲和 金属 匀浆
Defensive Responses of Rice Genotypes for Resistance Against Rice Leaffolder Cnaphalocrocis medinalis 预览
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作者 M. PUNITHAVALLI N. M. MUTHUKRISHNAN M. BALAJI RAJKUMA 《水稻科学:英文版》 2013年第5期363-370,共8页
The experiment was carried out to assess the reaction of different categories of rice genotypes viz.,resistant,susceptible,hybrid,scented,popular and wild in response to the infestation by rice leaffolder(RLF),Cnaphal... The experiment was carried out to assess the reaction of different categories of rice genotypes viz.,resistant,susceptible,hybrid,scented,popular and wild in response to the infestation by rice leaffolder(RLF),Cnaphalocrocis medinalis(Guenee)and to explore the possible use of these genotypes in developing RLF-resistant rice varieties.The changes of various biochemical constituents such as leaf soluble protein,phenol,ortho-dihydroxy phenol,tannin and enzymes viz.,peroxidase,phenyl alanine ammonia lyase(PAL)were assessed spectrophotometrically in all the rice genotypes before and after RLF infestation.The protein profile was analyzed using sodium dodecyl sulphate-poly acrylamide gel electrophoresis(SDS-PAGE)method.A significant constituent of biochemical content such as tannin,phenol and ortho-dihydroxy phenol has been increased along with enzyme activities of peroxidase and PAL in the infested resistant(Ptb 33,TKM6 and LFR831311)and wild rice genotypes(Oryza minuta and O.rhizomatis).A decrease in leaf protein content was evident invariably in all the infested rice genotypes.It is also evident that the contents of biochemicals such as phenol,orthodihydroxy phenol and tannin were negatively correlated with leaffolder damage.However,leaf protein content was positively correlated with the damage by rice leaffolder.SDS-PAGE analysis for total protein profiling of healthy and C.medinalis-infested genotypes revealed the enhanced expression of a high molecular weight(>97 kDa)protein in all the genotypes.Besides,there was also an increased induction of a 38 kDa protein in C.medinalis infested resistant genotypes,which was absent in uninfested plants.The present investigation proved that the elevated levels of biochemicals and enzymes may play a vital role in rice plants resistance to RLF. 展开更多
关键词 水稻基因型 稻纵卷叶螟 防御反应 SDS-PAGE分析 聚丙烯酰胺凝胶电泳法 小粒野生稻 苯丙氨酸解氨酶 十二烷基硫酸钠
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Purification of Antimicrobial Peptide from Antarctic Krill (Euphausia superba) and its Function Mechanism 预览 被引量:1
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作者 ZHAO Ling YIN Bangzhong +1 位作者 LIU Qi CAO Rong 《中国海洋大学学报:英文版》 SCIE CAS 2013年第3期484-490,共7页
The preliminary purification and antimicrobial mechanism of antimicrobial peptide from Antarctic Krill were studied in this paper.The results showed that the molecular weight range of antimicrobial polypeptide(CMCC-1)... The preliminary purification and antimicrobial mechanism of antimicrobial peptide from Antarctic Krill were studied in this paper.The results showed that the molecular weight range of antimicrobial polypeptide(CMCC-1) obtained by cation exchange chromatography was between 245-709D as detected by molecular sieve chromatography,and the minimum inhibition concentration(MIC) of CMCC-1 against Staphylococcus aureus was 5.0 mg mL-1.The antimicrobial mechanism of CMCC-1 was studied with S.aureus as indicator bacterium.Compared with control group,the results of the experimental group in which S.aureus was treated with CMCC-1 were as follows: 1) CMCC-1 could inhibit cell division at logarithmic phase.2) The protein and reducing sugar content,and the conductivity of culture medium increased,and the activity of alkaline phosphatase and β-galactosidase could be detected in the culture medium.3) Observation under scanning electron microscope revealed that somatic morphology became irregular,and then somatic surface became coarse.The cell became much smaller,and most somatic cells gathered.The boundary between cells became dim and finally fused as a whole.4) Observation under transmission electron microscope showed that the surface of S.aureus became rough and the reproducing ability was restrained.The cell wall became thin and the cytoplasm shrunk.Substances inside cell leaked out,which caused cells death.5) SDS-PAGE analysis showed that some bands disappeared,and the residual bands became vague.6) The genomic DNA electrophoresis results showed that the genomic DNA bands of S.aureus were not degraded but the brightness significantly reduced.Thus,it is supposed that CMCC-1 could destroy the cell wall and membrane of S.aureu,increase the cell membrane permeability and the leaking-out of intracellular substances,and thus cause the death of S.aureu. 展开更多
关键词 南极磷虾 抗菌机理 抗菌肽 金黄色葡萄球菌 纯化 SDS-PAGE分析 阳离子交换层析 基因组DNA
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猪、牛肉加热终点温度检测方法的研究 预览 被引量:3
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作者 刘伟 贾晓川 +3 位作者 赵良娟 贺艳 张宏伟 郑文杰 《食品研究与开发》 CAS 北大核心 2012年第6期共4页
为满足肉类及肉制品加热终点温度监控需要,通过蛋白质SDS-PAGE电泳来检测加热终点温度指示蛋白乳酸脱氢酶含量,通过肉产品中加热与否的LDH SDS-PAGE电泳LDH量的变化来进行加热终点温度的检测,结果表明,建立的SDS-PAGE加热终点温度检测... 为满足肉类及肉制品加热终点温度监控需要,通过蛋白质SDS-PAGE电泳来检测加热终点温度指示蛋白乳酸脱氢酶含量,通过肉产品中加热与否的LDH SDS-PAGE电泳LDH量的变化来进行加热终点温度的检测,结果表明,建立的SDS-PAGE加热终点温度检测方法可以有效的进行鉴别检测需要. 展开更多
关键词 猪牛肉 加热终点温度 SDS-PAGE分析
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Effect of Limited Enzymatic Hydrolysis on Structure Characterization of Glycinin 预览
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作者 Chunhong DUAN Wan SUN +2 位作者 Yunjie LI Yunlong WANG Siyi PAN 《农业生物技术:英文版》 2012年第3期52-54,共3页
[Objective] This study aimed to characterize the structure of soybean glycinin affected by limited enzymatic hydrolysis.[Method] The glycinin was limitedly hydrolyzed by alkaline protease;then the SDS-polyacrylamide g... [Objective] This study aimed to characterize the structure of soybean glycinin affected by limited enzymatic hydrolysis.[Method] The glycinin was limitedly hydrolyzed by alkaline protease;then the SDS-polyacrylamide gel electrophoresis(SDS-PAGE),scanning electron microscopy(SEM),Fourier transform infrared spectroscopy(FTIR) and other means were performed to characterize the glycinin structure changing during the hydrolysis process.[Result] SDS-PAGE analysis showed that the subunit content of glycinin significantly decreased after hydrolysis,and acidic subunits were more susceptible to hydrolysis than alkaline subunits.The scanning electron microscopy revealed that the structure of glycinin powder changed greatly after hydrolysis.The FTIR results showed that the proportions of all conformations of glycinin changed greatly during hydrolysis process.In addition,the protein hydrophobicity and sulfhydryl content were also significantly influenced by hydrolysis.[Conclusion] The enzymatic hydrolysis greatly changed the conformations of glycinin,and the change was dependent on the degree of hydrolysis. 展开更多
关键词 大豆球蛋白 结构表征 SDS-PAGE分析 SDS-聚丙烯酰胺凝胶电泳 有限酶解 电子显微镜观察 傅里叶变换红外光谱 碱性蛋白酶
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Cloning and characterization of a thermostable carboxylesterase from inshore hot spring thermophile Geobacillus sp. ZH1 预览
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作者 ZHU Yanbing LIU Guangming +3 位作者 LI Hebin LIU Jingwen BAI Xiaoming CAI Huinong 《海洋学报:英文版》 SCIE CAS CSCD 2012年第6期117-126,共10页
从 thermophilic 细菌 Geobacillus sp 编码 carboxylesterase 的基因(741 bp ) 。ZHl 被克隆并且在 Escherichia coli 的 overexpressed。净化的 recombinant 蛋白质由 SDS 页分析介绍了大约 40 kDa 的一个分子的团。当底层证实了它的 ... 从 thermophilic 细菌 Geobacillus sp 编码 carboxylesterase 的基因(741 bp ) 。ZHl 被克隆并且在 Escherichia coli 的 overexpressed。净化的 recombinant 蛋白质由 SDS 页分析介绍了大约 40 kDa 的一个分子的团。当底层证实了它的 esterase 活动,用有不同的酰的 p-nitrophenyl 酉旨的酶试金锁住长度,产出有 p-nitrophenyl 醋酸盐的最高特定的活动。在测试的 p-nitrophenyl 酉旨之中, carboxylesterase 更高效地为 p-nitrophenyl caprylate,而是 hydrolyzed p-nitrophenyl 丁酸盐介绍了偏爱。当 p-nitrophenyl 丁酸盐被用作底层时, recombinant carboxylesterase 在 pH 展出了最高的活动 8.0 和 60 瑡潩 ? 景挠汨牯灯票汬愠 ? 桃 ??? 慳楬楮祴 ? 楤獳汯敶 ? 湩牯慧楮 ? 楮牴杯湥 ???? 楤獳汯敶 ? 硯杹湥 ? 佄 ? 湡 ? 楳楬慣整 ? 楓 ?? 慷 ? 敲灳湯楳汢? 潦 ? 桴 ? 慶楲瑡潩獮椠 ? 潺灯慬歮潴 ? 潣浭湵瑩 ? 瑳畲瑣牵 ? 湩琠敨匠湡敭 ? 慂吗? 展开更多
关键词 嗜热芽孢杆菌 羧酸酯酶 耐热性 SDS-PAGE分析 克隆 对硝基苯基 酯酶活性 鉴定
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Occurrence and Characterization of Pale,Soft,Exudative-Like Broiler Muscle Commercially Produced in China 预览
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作者 ZHU Xue-shen XU Xing-lian MIN Hui-hui ZHOU Guang-hong 《农业科学学报:英文版》 SCIE CSCD 2012年第8期1384-1390,共7页
Pale, soft, exudative-like (PSE-like) broiler muscle is a growing problem for meat industry all over the world. However, limited studies have been made to assess broiler meat quality in China. The aim of this study wa... Pale, soft, exudative-like (PSE-like) broiler muscle is a growing problem for meat industry all over the world. However, limited studies have been made to assess broiler meat quality in China. The aim of this study was to investigate the characteristics and incidence of PSE-like broiler muscle commercially produced in China. A total of 1 274 Pectoralis muscles of Arbor Acre broiler were randomly obtained from the processing line to determine the commercial incidence of PSE-like muscle based on color. Furthermore, broiler Pectoralis muscles selected from the 1 274 muscle samples were classified as PSE-like muscle (L*>53, n=33) and normal muscle (L*>48 and L*=53, n=33) to assess meat quality. It was determined that PSE-like muscle had lower muscle pH values, lower water-holding capacity (WHC), lower sarcoplasmic protein solubility, and lower total protein solubility than the normal muscle did. SDS-PAGE profile also showed that bands of approximate 96 and 24 kDa in sarcoplasmic protein and myofibrillar protein varied between these two groups, suggesting partial denaturation of sarcoplasmic proteins and precipitation on myofillarments. Correlation analysis showed that L* values have significant correlation with WHC and protein solubility. Furthermore, the distribution of L* values exhibited a normal curve with range varying from 42.70 to 58.37. It was considered that approximately 23.39% of the population was PSE-like muscle. These results suggest that PSE-like meat can represent a significant portion of commercially processed broiler breast meat in China and that the L* value measurement could be used to sort broiler meat quality using a cut-off point. 展开更多
关键词 爱拔益加肉鸡 商业化生产 渗出性 肌肉 中国 柔软 SDS-PAGE分析 加工生产线
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Codon Optimization of SMAP.29 Gene and Its Expression in Pichia pastoris 预览
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作者 Yaojun REN Enpeng HE +1 位作者 Xinhua WANG Xinwen BO 《农业生物技术:英文版》 CAS 2012年第4期49-53,共5页
[Objective] This study aimed to optimize the codon of antimicrobial peptide SMAP-29 gene and express SMAP-29 in Pichia pastoris. [Method] According to the published amino acid sequence of SMAP-29, a gene encoding SMAP... [Objective] This study aimed to optimize the codon of antimicrobial peptide SMAP-29 gene and express SMAP-29 in Pichia pastoris. [Method] According to the published amino acid sequence of SMAP-29, a gene encoding SMAP-29 (sheep myeloid antibacterial peptides-29) mature peptide was synthesized and was biased codon usage of Pichia pastoris. The synthesized gene was inserted into Pichia pastoris expression vector pPIC3.5K, transformed into Pichia pastoris strain GS115 by electroporation, and screened with G418 for high-copy recombinants. The methanol utility plus phenotype (Mut+) was selected with MM, MD and PCR. The correctly constructed recombinant strain was induced with methanol. Expression of SMAP-29 was analyzed by lysing the induced yeast cells with Tricine-SDS-PAGE. [Result] After induced with methanol for 2 d, a 3.2 kD induced band was detected in the cell lysate which was consistent to the predicted molecular weight of SMAP-29; the expression products purified with gel filtration chromatography showed significant antibacterial effect against Staphyloccocus aureus and Monilia albican but no significant antibacterial effect against Escherichia coli. [Conclusion] This study laid the foundation for the application of SMAP-29 in biomedicine, agriculture and other fields.更多还原 展开更多
关键词 密码子优化 毕赤酵母 编码基因 SDS-PAGE分析 金黄色葡萄球菌 凝胶过滤色谱法 细胞裂解液 酵母表达载体
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