关键词: 镰形扇头蜱, 唾液腺, 抑制消减杂交, cDNA文库, 表达序列标签

Abstract: To screen and analyze the differentially expressed genes from salivary gland of Rhipicephalus haemaphysaloides haemaphysaloides, two salivary gland subtracted cDNA libraries of female half-feeding blood and male feeding blood ticks were constructed in the clone vector of pGEM-T-easy by the suppression subtractive hybridization (SSH) method, respectively. 247 female ESTs and 168 male ESTs were obtained from the two SSH cDNA libraries. There were 25 ESTs including 5’ end and 44 ESTs including 3’ end in the female ESTs and 53 ESTs including 5’ end and 74 ESTs including 3’ end in the male ESTs, respectively. To identify the subtraction efficiency of the two SSH cDNA libraries, RT-PCR method was used to test 24 female ESTs and 21 male ESTs selected randomly and not repeatedly. The results showed that there were 13 upregulated or new expressed genes in half-feeding blood salivary gland of the female Rhipicephalus haemaphysaloides and it revealed that the subtraction efficiency was 54%, and there were 9 upregulated or new expressed genes in feeding blood salivary gland of the male Rhipicephalus haemaphysaloides and it revealed that the subtraction efficiency was 43%, respectively. 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. In general, most of the proteins of the libraries were mainly related to the feeding blood physiology of the ticks including anticoagulation, immunity regulation, energy metabolism and transcription.

Key words: Rhipicephalus haemaphysaloides haemaphysaloides, Salivary gland, cDNA library, Expression sequence tag (EST), Suppression subtractive hybridization (SSH)