摘要
为明确全沟硬蜱涎腺Salp15蛋白家族的基因表达情况，本文根据已知的Salp15序列设计引物对全沟硬蜱幼蜱的总RNA进行RT—PCR，克隆到可与莱姆病螺旋体外膜蛋白OspC互作的涎腺蛋白Salp15CDS序列，命名为I．Plarva5。对该蛋白做生物信息学预测，其肽链在N端含有信号肽，在c端含有与CD4分子的结合位点。与其他蜱种Salp15蛋白的相似性分别为：62．3％（北美的肩突硬蜱Ixodesscapularis）、74．5％（欧洲的篦子硬蜱Lricinusiric3）、59．4％（西伯利亚全沟硬蜱Lpersulcatus）、67．6％（日本地区的全沟硬蜱iperl）。将该cDNA序列分别加上EcoRI，SalI酶切位点，插入pMAL．c4X表达载体上，经IPTG诱导。在Transetta（DE3）上融合蛋白成功表达。此工作为研究全沟硬蜱的Salp15与莱姆病螺旋体，以及与宿主动物的相互关系奠定了基础。
As one of main vectors for Lyme borreliosis, Ixodes persulcatus threats both innocent individuals and domestic livestock. Salpl5, a 15-kDa salivary protein in Ixodid ticks, has been known to suppress against the activities of host immunity and enhance the transmission of Lyme borrelia. To determine Salpl5-like gene in lxodes persulcatus, the primers for RT-PCR against total RNA from nymph according to other Salpl5 homologues were designed, and firstly the Salpl5 protein entitling I. p larva5 （ HE820731 ） potentially interacting with the outer surface protein C （ OspC ） of Lyme spirochaeta was obtained. The deduced protein I. p larva5 shows similarity to Salpl5 from other speices, i. e. 62. 3% （I. scapularis in North America）, 74. 5% （1. ricinus in Western Europe）, 59.4% （I. persulcatus in Siberia）, 67.6% （I. persulcatus in Japan） respectively. I. p larva5 CDS was then subcloned into expression vector pMAL-c4X, expressing in E. coll Rosseta （DE3）, induced by IPTG. The recombinant protein was confirmed expressed successfully by SDS-PAGE and western blot, and its relative molecular weights about 60 kDa with MBP Taq protein. This study lays the fundamental basis for the interacting between spirochaeta and Ixodes persulcatus and host reservoir.
出处
《寄生虫与医学昆虫学报》
CAS
2013年第2期102-109,共8页
Acta Parasitologica et Medica Entomologica Sinica
基金
国家自然科学基金资助项目（No.81271878
No.30872196）
关键词
涎腺蛋白
全沟硬蜱
宿主免疫
克隆
表达
Salp15
lxodes persulcatus
host immunity
clone
expression
