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

doi:10.1016/S1671-2927(00)8685

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (9): 1528-1536.DOI: 10.1016/S1671-2927(00)8685

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

XIANG Fei-yu, ZHOU Yong-zhi, ZHOU Jin-lin

Key Laboratory of Animal Parasitology, Ministry of Agriculture/Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, P.R.China

收稿日期:2011-06-06 出版日期:2012-09-01 发布日期:2012-09-09
通讯作者: Correspondence ZHOU Jin-lin, Tel: +86-21-34293411, Fax: +86-21-54081818, E-mail: jinlinzhou@hotmail.com
作者简介:XIANG Fei-yu, Mobile: 13996999752, E-mail: geneclub@hotmail.com
基金资助:
This work was supported by the National Natural Science Foundation of China (31172095).

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

XIANG Fei-yu, ZHOU Yong-zhi, ZHOU Jin-lin

Key Laboratory of Animal Parasitology, Ministry of Agriculture/Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, P.R.China

Received:2011-06-06 Online:2012-09-01 Published:2012-09-09
Contact: Correspondence ZHOU Jin-lin, Tel: +86-21-34293411, Fax: +86-21-54081818, E-mail: jinlinzhou@hotmail.com
About author:XIANG Fei-yu, Mobile: 13996999752, E-mail: geneclub@hotmail.com
Supported by:
This work was supported by the National Natural Science Foundation of China (31172095).

摘要/Abstract

摘要： 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)

Abstract: 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.

Key words: Rhipicephalus haemaphysaloides, salivary gland, suppression subtractive hybridization (SSH), cDNA library, expression sequence tag (EST)

XIANG Fei-yu, ZHOU Yong-zhi, ZHOU Jin-lin. Identification of Differentially Expressed Genes in the Salivary Gand of Rhipicephalus haemaphysaloides by the Suppression Subtractive Hybridization Approach[J]. Journal of Integrative Agriculture, 2012, 12(9): 1528-1536.

参考文献

[1]Bhat H R, Naik S V, Ilkal M A, Banerjee K. 1978. Transmission of Kyasanur Forest disease virus by Rhipicephalus haemaphysaloides ticks. Acta Virology, 22, 241-244.

[2]Bior A D, Essenberg R C, Sauer J R. 2002. Comparison of differentially expressed genes in the salivary glands of male ticks, Amblyomma americanum and Dermacentor andersoni. Insect Biochemisty and Molecular Biology, 32, 645-655.

[3]Bowman A S, Coons L B, Needham G R, Sauer J R. 1997. Tick saliva: recent advances and implications for vector competence. Medical and Veterinary Entomology, 11, 277-285.

[4]Diatchenko L, Lau Y F, Campbell A P, Chenchik A, Moqadam F, Huang B, Lukyanov S, Lukyanov K, Gurskaya N, Sverdlov E D, et al. 1996. Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. Proceedings of the National Academy of Sciences of the United States of America, 93, 6025-6030.

[5]Feldman-Muhsam B, Borut S, Saliternik-Givant S. 1970. Salivary secretion of the male tick during copulation. Journal of Insect Physiology, 16, 1945-1949.

[6]Francischetti I M, Valenzuela J G, Andersen J F, Mather T N, Ribeiro J M. 2002. Ixolaris, a novel recombinant tissue factor pathway inhibitor (TFPI) from the salivary gland of the tick, Ixodes scapularis: identification of factor X and factor Xa as scaffolds for the inhibition of factor VIIa/tissue factor complex. Blood, 99, 3602-3612.

[7]Food and Agricultural Organization. 1984. Ticks and Tickborne Disease Control: A Practical Field Manual. vol. 1. Food and Agricultural Organization, Rome. pp. iv-xi.

[8]Hannier S, Liversidge J, Sternberg J M, Bowman A S. 2003. Ixodes ricinus tick salivary gland extract inhibits IL-10 secretion and CD69 expression by mitogen-stimulated murine splenocytes and induces hyporesponsiveness in B lymphocytes. Parasite Immunology, 25, 27-37.

[9]Leboulle G, Crippa M, Decrem Y, Mejri N, Brossard M, Bollen A, Godfroid E. 2002. Characterization of a novel salivary immunosuppressive protein from Ixodes ricinus ticks. The Journal of Biological Chemistry, 277, 10083-10089.

[10]Leboulle G, Rochez C, Louahed J, Ruti B, Brossard M, Bollen A, Godfroid E. 2000. Isolation of Ixodes ricinus salivary gland mRNA encoding factors induced during blood feeding. American Journal of Tropical Medicine and Hygiene, 66, 225-233.

[11]Nakajima C, da Silva Vaz I J, Imamura S, Konnai S, Ohashi K, Onuma M. 2005. Random sequencing of cDNA library derived from partially-fed adult female Haemaphysalis longicornis salivary gland. The Journal of Veterinary Medical Science, 67, 1127-1131.

[12]Nuttall P A, Trimnell A R, Kazimirova M, Labuda M. 2006. Exposed and concealed antigens as vaccine targets for controlling ticks and tick-borne diseases. Parasite Immunology, 28, 155-163.

[13]Oaks J F, McSwain J L, Bantle J A, Essenberg R C, Sauer J R. 1991. Putative new expression of genes in ixodid tick salivary gland development during feeding. Journal of Parasitology, 77, 378-383.

[14]Puente X S, Sanchez L M, Overall C M, Lopez-Otin C. 2003. Human and mouse proteases: a comparative genomic approach. Nature Reviews Genetics, 4, 544-558.

[15]Ribeiro J M, Alarcon-Chaidez F, Francischetti I M, Mans B J, Mather T N, Valenzuela J G,Wikel S K. 2006. An annotated catalog of salivary gland transcripts from Ixodes scapularis ticks. Insect Biochemistry and Molecular Biology, 36, 111-129.

[16]Sangamnatdej S, Paesen G C, Slovak M, Nuttall P A. 2002. A high affinity serotonin-and histamine-binding lipocalin from tick saliva. Insect Molecular Biology, 11, 79-86.

[17]Sauer J R, Essenberg R C, Bowman A S. 2000. Salivary glands in ixodid ticks:control and mechanism of secretion. Journal of Insect Physiology, 46, 1069-1078.

[18]Shipley M, Dillwith J, Bowman A, Essenberg R, Sauer J. 1993. Changes in lipids from the salivary glands of the lone star tick, Amblyomma americanum, during feeding. Journal of Parasitology, 79, 834-842.

[19]Valenzuela J G, Francischetti I M, Pham V M, Garfield M K, Mather T N, Ribeiro J M. 2002. Exploring the sialome of the tick, Ixodes scapularis. Journal of Experimental Biology, 205, 2843-2864.

[20]Valenzuela J G, Francischetti I M, Pham V M, Garfield M K, Mather T N, Ribeiro J M. 2002. Exploring the sialome of the tick, Ixodes scapularis. Journal of Experimental Biology, 205, 2843-2864.

[21]Wang H, Nuttall P A. 1994. Excretion of host immunoglobulin in tick saliva and detection of IgG-binding proteins in tick haemolymph and salivary glands. Parasitology, 109, 525-530.

[22]Wang H, Nuttall P A. 1995. Immunoglobulin G binding proteins in the ixodid ticks, Rhipicephalus appendiculatus, Amblyomma variegatum and Ixodes hexagonus. Parasitology, 111, 161-165.

[23]Wang H, Nuttall P A. 1999. Immunoglobulin-binding proteins in ticks: new target for vaccine development against a blood-feeding parasite. Cellular and Molecular Life Sciences, 56, 286-295.

[24]Wang H, Paesan G C, Nuttall P A, Barbour A G. 1998. Male ticks help their mates to feed. Nature, 391, 753-754.

[25]Waxman L, Smith D E, Arcuri K E, Vlasuk G P. 1990. Tick anticoagulant peptide (TAP) is a novel inhibitor of blood coagulation factor Xa. Science, 248, 593-596.

[26]Willadsen P, Smith D, Cobon G, McKenna R V. 1996. Comparative vaccination of cattle against Boophilus microplus with recombinant antigen Bm86 alone or in combination with recombinant Bm91. Parasite Immunology, 18, 241-246.

[27]Yin H, Lu W, Luo J. 1997. Babesiosis in China. Tropical Animal Health and Production, 29(Suppl. 4), 11S-15S.

[28]Zhou J, Gong H, Zhou Y, Xuan X, Fujisaki K. 2006. Identification of a glycine-rich protein from the tick Rhipicephalus haemaphysaloides and evaluation of its vaccine potential against tick feeding. Parasitology Research, 100, 77-84.

[29]Zhou Y Z, Zhou J L, Cao J, Gong H Y. 2003. A laboratory method for the maintaining and feeding of four ixodid ticks and observation on their partial biology characters. Chinese Journal of Veterinary Parasitology, 11, 25-27.

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

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

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 5

编辑推荐

Metrics

[1]	ZHANG Xian-wei, LI Zi-cong, MENG Fan-ming, WANG De-hua, LIU De-wu, HE Xiao-yan, SUN Yue, BAI Yin-shan, WU Zhen-fang. Characterization of dual enzyme resulted from bicistronic expression of two β-glucanases in porcine cells[J]. Journal of Integrative Agriculture, 2015, 14(4): 732-740.
[2]	ZHAO Su-mei, LIU Yong-gang, PAN Hong-bing, ZHANG Xi, GE Chang-rong, JIA Jun-jing , GAO Shi-zheng. Generation and Analysis of Expressed Sequence Tags (ESTs) from Muscle Full-Length cDNA Library of Wujin Pig[J]. Journal of Integrative Agriculture, 2014, 13(2): 378-386.
[3]	LI Liang, WANG Wei-qi, HAN Tian-fu, HOU Wen-sheng. Construction of Two Suppression Subtractive Hybridization Libraries and Identification of Salt-Induced Genes in Soybean[J]. Journal of Integrative Agriculture, 2012, 12(7): 1075-1085.
[4]	PANG Rui-hua, MA Guang-ying, LOU Xue-yuan, BAO Man-zhu , YE Yao-mei. Construction and Analysis of Suppression Subtractive cDNA Library from Fertile Disk Floret Buds of Zinnia elegans [J]. Journal of Integrative Agriculture, 2012, 12(4): 567-575.
[5]	KE Tao; DONG Cai-hua; MAO Han; ZHAO Ying-zhong; LIU Hong-yan and LIU Sheng-yi. Construction of a Normalized Full-Length cDNA Library of Sesame Developing Seed by DSN and SMART[J]. Journal of Integrative Agriculture, 2011, 10(7): 1004-1009.