Molecular characterization and protective efficacy of silent information regulator 2A from Eimeria tenella.
Molecular characterization and protective efficacy of silent information regulator 2A from Eimeria tenella.
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柔嫩艾美耳球虫沉默信息调节因子 2A 的分子表征和保护功效
DOI：
10.1186/s13071-016-1871-0
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发表时间：
2016-11-25
影响因子：
3.2
通讯作者：
Huang B
中科院分区：
文献类型：
--
作者：
Dong H;Yang S;Zhao Q;Han H;Zhu S;Zhu X;Li C;Wang Z;Xia W;Men Q;Yang L;Huang B
沉默信息调节因子2（SIR 2）蛋白是NAD +依赖性蛋白脱乙酰酶家族，被认为是抗寄生虫剂的潜在靶标。在这项研究中，我们克隆和表征SIR 2A的原生动物寄生虫柔嫩艾美耳球虫（EtSIR 2A），并研究其作为DNA疫苗的保护效力。EtSIR 2A基因编码33.37kDa的蛋白质。克隆了tenella第二代裂殖子，并在大肠杆菌表达系统中产生重组EtSIR 2A蛋白（rEtSIR 2A）。rEtSIR 2A用于免疫家兔。抗rEtSIR 2A抗体用于通过免疫荧光测定（IFA）确定寄生虫中EtSIR 2A的免疫定位。EtSIR 2A在大肠杆菌不同发育阶段的转录和蛋白表达。分别通过实时定量PCR（qPCR）和蛋白质印迹（WB）分析来观察tenella。构建了重组质粒pCAGGS-EtSIR 2A，并对其抗大肠杆菌的效果进行了初步研究。评估鸡的柔嫩体感染。qPCR和WB分析显示EtSIR 2A的表达在mRNA和蛋白质水平上都受到发育调节。EtSIR 2A mRNA水平在未形成孢子的卵囊中高于其他发育阶段，包括形成孢子的卵囊、子孢子和第二代裂殖子。相反，EtSIR 2A蛋白表达水平在第二代裂殖子中最高，在未形成孢子的卵囊和形成孢子的卵囊中中等，在子孢子中最低。抗rEtSIR 2A抗体的免疫组化结果表明，EtSIR 2A主要定位于子孢子和第二代裂殖子的胞质中，在第一阶段裂殖子中强烈表达。动物攻毒实验表明，pCAGGS-EtSIR 2A免疫鸡显著增加平均体重增加，并降低平均病变评分和卵囊产量。这些结果表明，EtSIR 2A可能在寄生虫细胞存活中起重要作用，并可能成为开发抗E.鸡的柔嫩杆菌感染。
Silent information regulator 2 (SIR2) proteins are a family of NAD + -dependent protein deacetylases that are considered potential targets for anti-parasitic agents. In this study, we cloned and characterized SIR2A of the protozoan parasite Eimeria tenella (EtSIR2A) and investigated its protective efficacy as a DNA vaccine. The EtSIR2A gene encoding 33.37 kDa protein from E. tenella second-generation merozoites was cloned, and recombinant EtSIR2A protein (rEtSIR2A) was produced in an Escherichia coli expression system. The rEtSIR2A was used to immunize rabbits. Anti-rEtSIR2A antibodies were used to determine the immunolocolization of EtSIR2A in the parasite by immunofluorescence assay (IFA). Transcript and protein expression of EtSIR2A in different development stages of E. tenella were observed by quantitative real-time PCR (qPCR) and western blot (WB) analysis, respectively. The recombinant plasmid pCAGGS-EtSIR2A was constructed and its efficacy against E. tenella infection in chickens was evaluated. qPCR and WB analysis revealed EtSIR2A expression was developmentally regulated at both the mRNA and protein levels. EtSIR2A mRNA levels were higher in unsporulated oocysts than at other developmental stages, including sporulated oocysts, sporozoites and second-generation merozoites. In contrast, EtSIR2A protein expression levels were highest in second-generation merozoites, moderate in unsporulated oocysts and sporulated oocysts and lowest in sporozoites. Immunostaining with anti-rEtSIR2A antibody indicated that EtSIR2A was mainly located in the cytoplasm of sporozoites and second-generation merozoites, and was strongly expressed during first stage schizogony. Animal-challenge experiments demonstrated that immunization with pCAGGS-EtSIR2A significantly increased average body-weight gain, and decreased mean lesion score and oocyst output in chickens. These results suggest that EtSIR2A may play an important role in parasite cell survival and may be an effective candidate for the development of new vaccines against E. tenella infection in chickens.
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影响因子：
10.5
作者：
BRACHMANN, CB;SHERMAN, JM;BOEKE, JD
通讯作者：
BOEKE, JD
DOI：
10.1006/bbrc.2000.3000
发表时间：
2000-07-05
影响因子：
3.1
作者：
Frye, RA
通讯作者：
Frye, RA
影响因子：
11.4
作者：
García-Salcedo, JA;Gijón, P;Pays, E
通讯作者：
Pays, E
影响因子：
2
作者：
Jiang, Lianlian;Lin, Jiaojiao;Huang, Bing
通讯作者：
Huang, Bing
影响因子：
3.8
作者：
Lancelot J;Caby S;Dubois-Abdesselem F;Vanderstraete M;Trolet J;Oliveira G;Bracher F;Jung M;Pierce RJ
通讯作者：
Pierce RJ