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Study on Differentially Expressed Genes in the Salivary Gland of Hyalomma Asiaticum Tick

Author: ChengTianYin
Tutor: ShiQiShun;LinJiaoJiao
School: Hunan Agricultural University
Course: Animal Genetic Breeding and Reproduction
Keywords: Asian Hyalomma salivary glands Differentially expressed gene library GP29 gene GP15 gene Clone Expression
CLC: Q969.91
Type: PhD thesis
Year: 2005
Downloads: 105
Quote: 2
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Abstract


Selected from a single female ticks breeding herd is not blood-sucking female adult ticks 60, were randomly divided into two groups, a group of half-blood meal, and the other group do not suck blood. Separation is not vampire and half full of blood state tick salivary glands. After total RNA extraction, first-strand synthesis, long-distance amplification, Ras I digestion, add joint, net of hybridization and selective amplification step, the isolated cDNA fragment of the differentially expressed genes. The pure product of the second amplification product into pGEM T-Easy vector and transformed into DH5a competent. 120 fragment identified by cloning and sequencing, and 84 of them identified as expressed sequence tags (EST). RT-PCR analysis confirmed that the library randomly selected 10 fragments in the blood-sucking female adult ticks salivary gland expression, not blood-sucking expression. Sequence alignment with the fragment of the NCBI database, the results were as follows: 120 fragments in the library, 90 to find a sequence having a certain similarity, wherein the similarity with other known genes ticks fragment 21, and press mosquito, Culex pipiens, hookworm, Ostertagia and other blood-sucking parasite gene fragments have a certain similarity 19. The ticks known genes Similar 21 fragment, the fragment is identified as EST 5 wherein the poly (A), 2 having ACGCGGGG; 2 is both Poly (A) with ACGCGGGG (that is, full-length gene, one of which has been receiving Genbank accession number DQ021902). The HAB1 and Ha7.7 similarity EST screened from the ticks known gene. According to HAB1 sequence design amplification primers HAB1 unknown end HAB1-GSP 1 and HAB1 GSP 2 . With HAB1-GSP 1 and UAP amplified by half full blood female adult tick salivary gland total RNA was reverse transcribed into the first strand, HaB1-GSP 2 and AUAP nested amplification, that was HAB1 unknown 3'-end. According to HAB1 its 3'-terminal sequence splicing results, design to GP 29 of the full-length amplification primers HAB1-FLP , HAB1-FLP - , in order to amplify the cDNA first strand the new gene GP 29 (accession number: AY803896) full-length. As design amplified Ha7.7 the 5'-end of the primer Ha7.7-GSP 1 , Ha7.7-GSP 2 , Ha7.7-GSP 3 and amplified GP 15 of the full-length primer Ha7.7 the for-FLP , Ha7.7-FLP - . The clone Ha7.7 obtaining the unknown part and full-length GP 15 (accession number: DQ020265). RT-PCR analysis showed that: Asian Hyalomma tick salivary glands only two genes expressed in the blood-sucking.

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CLC: > Biological Sciences > Entomology > Applied Entomology (Economic Entomology ) > Ticks and mites SCIENCE
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