棒狀體相關蛋白1（rhoptry-associated prote她妹in 1，RAP-1）是由棒狀體分泌的吧笑與侵襲宿主細胞相關的蛋白。本研究以頂複門原蟲RAP-1基因爲靶遠腦标，用最大似然法構建了巴貝斯屬、瘧原蟲屬錢行、泰勒蟲屬的RAP-1基因進(jìn)化樹；應用軟件預測分析白明RAP-1蛋白的理化性質、親疏水性、跨膜區以及二、三級結構，并表達純化了馬姐筆泰勒蟲（Theileria equi）RAP-1重組蛋白。結果顯示：可機巴貝斯屬不同蟲株相聚類，測定序列與馬泰勒蟲聚個森爲一支，又與惡性瘧原蟲相聚類，表明親緣關系較近。馬泰勒蟲RAP老水-1蛋白理論等電點爲9.85，半衰期爲30 h線資，總平均親水性（GRAVY）爲-0.36樹得8，無跨膜區；RAP-1蛋白二級結構中，α數車-螺旋、β-折疊、β-轉角和無規卷曲占比分别們窗爲38.5%、15.5%、27.8%、18.1%；構建的三級人相結構立體展現了RAP-1蛋白形态。成(c光坐héng)功構建了原核表達載體pET-32a-RAP-1；該重組蛋白地開主要以包涵體形式存在，蛋白大小爲65 kDa。RAP-1蛋白可通舞都過(guò)原核表達系統高效表達，純化後(hò懂員u)的産物能(néng)被(bèi)馬泰勒蟲标準陽性血清識别，具有錢們良好(hǎo)的反應原性。本研究爲深妹商入了解馬泰勒蟲入侵宿主的機制機理以及RAP-1蛋白的功能(néng)研究司的奠定了基礎。
Prokaryotic Expression of RAP-1 Gene of員鐵 Theileriaequi and
Characteristic Analysis on the 麗開Encoded Protein
Rhoptry-associat也相ed protein 1（RAP-1）is a pro用子tein related to invading到畫 the host cells and secreted by 看筆the rhoptry. Target低呢ing at the RAP-1 gene of Protozoan綠但 protozoa，the gene evolut學爸ion trees of Babesia，Plasmod購雨ium and Theileria were 好關constructed by the maximum likelihood腦美 method. The physical and chemical prop一科erties，hydrophobicity，transmembr討化ane zone，secondary and tertiary struc務司tures of RAP-1 protein were河劇 predicted by software，and the r雪火ecombinant RAP-1 protein of Theileria通銀 equi was expressed and purifi那費ed. The results showed that the g業女enus of Babesia wa民生s clustered toget訊師her，its sequence and 腦熱that of Theileria e很腦qui belonged to the sam你裡e branch and clustered with the ge但他nus of Plasmodium falciparum，窗錢indicating that their ge都空nus was closely related. 熱你The theoretical isoelectric poi體吧nt of RAP-1 protein was 9.85，the西愛 half-life was 30 h理歌，the total average hydr工在ophilicity was -0.368，and there was no我文 transmembrane zone.明器 The secondary struc林飛ture α-helices，β-sheets，β-tu都老rns and random coils of RAP煙現-1 protein acco個匠unted for 38.5%，15.5%，27.8% and 18話喝.1%，respectively. The form of RAP-1 p快謝rotein was shown by the t年木hree-level structure，and th笑刀e prokaryotic expression vector pE請銀T-32a-RAP-1 was succ靜計essfully constructe也書d，the recombinant p校店rotein mainly existed in the fo有朋rm of inclusion body wit線章h the size of 65 kDa. In conclusio一個n，the RAP-1 protei冷在n could be highly expre紙靜ssed by prokaryo歌志tic expression system，and the purified湖市 products could be identified by the 國區standard positive serum of外們 Theileriaequi due to its go都靜od reactogenicity. The了民refore，a foundation was provided for 文拿further studying the mechanism of制問 Theileriaequi invading i話這nto hosts and the街都 functions of RA刀藍P-1 protein.
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