Nene Vishvanath, Lee Dan, Kang'a Simon, Skilton Robert, Shah Trushar, de Villiers Etienne, Mwaura Stephen, Taylor David, Quackenbush John, Bishop Richard
Parasite Genomics Department, The Institute for Genomic Research, 9712 Medical Center Drive, Rockville, MD 20850, USA.
Insect Biochem Mol Biol. 2004 Oct;34(10):1117-28. doi: 10.1016/j.ibmb.2004.07.002.
We describe the generation of an auto-annotated index of genes that are expressed in the salivary glands of four-day fed female adult Rhipicephalus appendiculatus ticks. A total of 9162 EST sequences were derived from an uninfected tick cDNA library and 9844 ESTs were from a cDNA library from ticks infected with Theileria parva, which develop in type III salivary gland acini. There were no major differences between abundantly expressed ESTs from the two cDNA libraries, although there was evidence for an up-regulation in the expression of some glycine-rich proteins in infected salivary glands. Gene ontology terms were also assigned to sequences in the index and those with potential enzyme function were linked to the Kyoto encyclopedia of genes and genomes database, allowing reconstruction of metabolic pathways. Several genes code for previously characterized tick proteins such as receptors for myokinin or ecdysteroid and an immunosuppressive protein. cDNAs coding for homologs of heme-lipoproteins which are major components of tick hemolymph were identified by searching the database with published N-terminal peptide sequence data derived from biochemically purified Boophilus microplus proteins. The EST data will be a useful resource for construction of microarrays to probe vector biology, vector-host and vector-pathogen interactions and to underpin gene identification via proteomics approaches.
我们描述了在饱血4天的成年雌性微小牛蜱唾液腺中表达的基因自动注释索引的生成。总共9162条EST序列来自未感染蜱的cDNA文库,9844条EST来自感染了泰勒虫的蜱的cDNA文库,泰勒虫在III型唾液腺腺泡中发育。两个cDNA文库中高表达的EST之间没有重大差异,尽管有证据表明感染唾液腺中一些富含甘氨酸的蛋白质表达上调。基因本体术语也被分配到索引中的序列,具有潜在酶功能的序列与京都基因与基因组百科全书数据库相连,从而能够重建代谢途径。有几个基因编码先前已鉴定的蜱蛋白,如肌动蛋白或蜕皮甾类受体以及一种免疫抑制蛋白。通过用从生化纯化的微小牛蜱蛋白获得的已发表的N端肽序列数据搜索数据库,鉴定出编码蜱血淋巴主要成分血红素脂蛋白同源物的cDNA。EST数据将成为构建微阵列的有用资源,用于探究媒介生物学、媒介-宿主和媒介-病原体相互作用,并通过蛋白质组学方法支持基因鉴定。