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Identification of Differentially Expressed Genes in the Salivary Gand of rhipicephalus haemaphysaloides by the Suppression Subtractive Hybridization Approach

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Journal of Integrative Agriculture 2012年第9期11卷 1528-1544页

作者： XIANG Fei-yu ZHOU Yong-zhi ZHOU Jin-lin Key Laboratory of Animal Parasitology Ministry of Agriculture~Shanghai Veterinary Research Institute Chinese Academy of AgriculturalSciences Shanghai 200241 P.R.China

For the purpose of screening and analyzing the differentially expressed genes from the salivary gland of rhipicephalus haemaphysaloides, two salivary gland-subtracted cDNA libraries of partially fed female ticks and fed male ticks were constructed using suppression subtractive hybridization （SSH）. A total of 247 female expression sequence tags （ESTs） and 168 male ESTs were obtained from the two SSH cDNA libraries. It is predicted that 25 female ESTs and 44 female ESTs contain the 5＂ and 3＂ ends, respectively, and that 53 male ESTs and 74 male ESTs contain the 5＂ and 3＂ ends, respectively. To identify the subtraction rate of the two SSH cDNA libraries, the RT-PCR method was used to test 24 female ESTs and 21 male ESTs selected randomly but not repeatedly. The results showed that there were 13 upregulated or differentially expressed genes in the partially fed salivary gland of the female R. haemaphysaloides and that the differentially expressed rate was 54%. In addition, they indicated that there were 9 upregulated or differently expressed genes in the fed salivary gland of the male R. haemaphysaloides and that the differentially expressed rate was 43%. Putative translations of 141 （57%） female ESTs and 125 （74%） male ESTs had similarity to GenBank sequences, and 32 （23%） female ESTs and 29 （23%） male ESTs exhibited similarity to tick proteins, which showed that most of the proteins in the libraries were mainly related to the feeding blood physiology of the ticks.

关键词： rhipicephalus haemaphysaloides salivary gland suppression subtractive hybridization (SSH) cDNA library expression sequence tag (EST)

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Proteomic Analysis of Proteins in the Salivary Glands of the Fed and Unfed Female Tick rhipicephalus haemaphysaloides

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Agricultural Sciences in China 2009年第1期8卷 121-127页

作者： XIANG Fei-yu ZHANG Jian-wu ZHOU Yong-zhi LI Zhuang GONG Hai-yan ZHOU Jin-lin Key Lab of Animal Parasitology of Ministry of Agriculture of China~Shanghai Veterinary Research Institute Chinese Academy of Agricultural Sciences Shanghai 200232 P.R.China Shanghai lnsititute of Cardiovascular Diseases Zhongshan Hospital Fudan University Shanghai 200032 P.R.China

Identification of differentially expressed salivary gland proteins between the fed and unfed female rhipicephalus haemaphysaloides may obtain valuable functional molecules. In this study, comparative two-dimensional gel electrophoresis （2-DE） and mass spectrometry were used to separate and identify differentially expressed salivary gland proteins between the fed and unfed female R. haemaphysaloides. The soluble proteins from the salivary glands of fed and unfed female R. haemaphysaloides were separated by a sequential extraction method followed by 2-DE and 2-DE images. Image analysis of the gels revealed 1 096 ± 87 protein spots from the fed female ticks and 991 ±64 protein spots from the unfed female ticks. Among those protein spots, about 724 ±34 were present both in the fed and unfed female ticks. Fourteen spots from the fed ticks and six spots from the unfed ticks were selected for peptide mass fingerprinting （PMF） and sequencing assay by mass spectrometry （MS）. Bioinformatic analysis showed that a majority of the differentially expressed proteins were involved in signal transduction, metabolism, and transcriptional regulation. These differentially expressed proteins might be antigen candidates for the development of vaccines against the tick.

关键词： rhipicephalus haemaphysaloides salivary g/and differentially expressed proteins two-dimensional gel electrophoresis mass spectrometry

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维普期刊数据库

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RNA interference and the vaccine effect of subolesin homolog from the tick rhipicephalus haemaphysaloides

RNA interference and the vaccine effect of subolesin homolog...

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中国畜牧兽医学会兽医寄生虫学分会第十三次学术研讨会

作者： Pengyun Lu Jie Cao Yingfang Yu Yongzhi Zhou Houshuang Zhang Haiyan Gong Guoqing Li Jinlin Zhou Key Laboratory of Animal Parasitology of Ministry of Agriculture Shanghai Veterinary Research InstituteChinese Academy of Agricultural Sciences College of Veterinary Medicine South China Agriculture University

Subolesin is a well-characterized protective antigen in many ticks,and thus,is potentially useful in the development of a broad-spectrum vaccine or autocidal control strategy using a gene knockdown to control tick infestations.A subolesin homolog was cloned from the Chinese dominant tick rhipicephalus haemaphysaloides by *** full length cDNA was 1,386 bp long,containing an ORF of 483 bp with a predicted molecular mass of 18.7 kDa and an isoelectric point of *** subolesin protein had a typical nuclear localization signal(NLS) signal at its N-terminal *** the full length cDNA of *** showed 52%and 80%identities with Ixodes scapularis and ***,respectively,the amino acid sequence alignments showed 80%and 97%identities,*** native subolesin,5kDa,was recognized in an unfed tick midgut using an antibody against recombinant subolesin.A transcriptional analysis showed that subolesin was expressed in the tick’s four developmental stages and in all of the tissues examined,except for synganglion.A real-time PCR analysis showed that the subolesin transcript was induced by Babesia microti,not by bacterial *** gene silencing in larvae and nymph ticks by RNAi inhibits blood feeding,but not ***,vaccinating with the recombinant subolesin induced a significant protective effect,resulting in the reduction of blood feeding and *** results encourage further studies using subolesin for the control of tick infestations in China.

关键词： rhipicephalus haemaphysaloides subolesin RNAi vaccine

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