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Anti-encystment and amoebicidal activity of Lonicera japonica Thunb. and its major constituent Chlorogenic acid in vitro 被引量:2
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作者 Tooba Mahboob Abdul-Majid Azlan +4 位作者 Tian-Chye Tan Chandramathi Samudi Shamala Devi Sekaran Veeranoot Nissapatorn Christophe Wiart 《Asian Pacific Journal of Tropical Medicine》 SCIE CAS 2016年第9期844-850,共7页
Objective:To examine the acanthamoebicidal effects of ethyl acetate,aqueous and butanol fractions of dried flower buds of Lonicera japonica(L.japonica) Thunb.(Flos Lonicerae) in vitro.Methods:Acanthamoeba triangularis... Objective:To examine the acanthamoebicidal effects of ethyl acetate,aqueous and butanol fractions of dried flower buds of Lonicera japonica(L.japonica) Thunb.(Flos Lonicerae) in vitro.Methods:Acanthamoeba triangularis isolates were obtained from environmental water samples and identified by PCR.They were exposed to ethyl acetate,water and butanol fractions of L.japonica Thunb.at concentrations ranging from 0.5 mg/m L to 1.5 mg/m L.The extracts were evaluated for growth inhibition at 24,48 and 72 h,respectively.Chlorogenic acid at a concentration of 1 mg/mL was examined for inhibition of encystment.Results:Ethyl acetate fraction at a concentration of 1.5 mg/mL evoked a significant reduction of trophozoite viability by 48.9% after 24 h,49.2% after 48 h and 33.7% after 72 h Chlorogenic acid,the major active constituent of L.japonica Thunb.at the concentration of 1 mg/mL reduced the cysts/trophozoite ratio by 100% after 24 h,84.0% after 48 h and 72.3% after 72 h.This phenolic compound at concentration of 1 mg/mL concurrent with 0.6% hydrogen peroxide inhibited hydrogen peroxide-induced encystment by 92.8% at 72 h.Conclusions:Results obtained from this study show that ethyl acetate fraction at 1.5 mg/m L is the most potent fraction of L.japonica Thunb.and its major constituent chlorogenic acid showed the remarkable inhibition of encystment at a concentration of 1 mg/mL. 展开更多
关键词 acanthamoeba spp. Amoebicidal Chlorogenic acid ENCYSTMENT LONICERA japonica Thunb.
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棘阿米巴原虫actin1基因原核表达载体构建与鉴定 被引量:1
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作者 李垚艳 梁宸 +3 位作者 杨春梅 王双露 李金旺 冯宪敏 《中国兽医学报》 CAS CSCD 北大核心 2018年第9期1745-1747,1752,共4页
棘阿米巴角膜炎的临床诊断比较困难,以角膜刮片的病原学诊断为主,样本采集困难,临床诊断率低。本研究构建了棘阿米巴原虫actin1基因进行原核表达载体,并进行鉴定。以棘阿米巴滋养体cDNA为模板,通过特异性引物,经聚合酶链反应,获得actin... 棘阿米巴角膜炎的临床诊断比较困难,以角膜刮片的病原学诊断为主,样本采集困难,临床诊断率低。本研究构建了棘阿米巴原虫actin1基因进行原核表达载体,并进行鉴定。以棘阿米巴滋养体cDNA为模板,通过特异性引物,经聚合酶链反应,获得actin1基因开放阅读框(ORF)片段,进行克隆及序列分析,经双酶切,连接,构建重组表达载体pET22b(+)-actin1。将重组载体转化入大肠杆菌(E.coli)DH5α,筛选出阳性菌落,经PCR扩增和酶切鉴定后,阳性质粒送样测序;提取阳性质粒,转化入E.coli BL21(DE+),1mol/L IPTG诱导重组蛋白r-actin1的表达。自棘阿米巴滋养体cDNA扩增得到约774bp的actin1基因片段;经PCR和酶切鉴定表明,重组表达载体pET22b(+)-actin1构建成功;重组菌株BL21(DE+)/pET22b(+)-actin1,经培养、诱导、SDS-PAGE电泳,出现1条相对分子质量约为30 000重组蛋白条带,r-actin1表达诱导表达成功。成功构建重组表达载体pET22b(+)-actin1,并诱导表达。 展开更多
关键词 棘阿米巴 肌动蛋白-1 重组载体 原核表达
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