CAJ | 학술논문

本研究对亚洲璃眼蜱多肽Ha-11的编码基因进行了克隆、表达和初步的功能分析。该基因开放阅读框（open reading frame, ORF）为372 bp，编码123个氨基酸，其中含23个氨基酸的信号肽。将去除信号肽的编码区基因亚克隆至pGEX-4T-1载体，大肠杆菌中诱导表达，获得了37 kDa重组蛋白。抗重组Ha-11蛋白的免疫血清识别半饱血雌蜱的唾液腺中约11 kDa的天然蛋白。Real-time PCR结果表明该基因在亚洲璃眼蜱的各发育阶段、各组织中均有表达，但以若蜱表达量最高，而组织中以淋巴液表达量最高。重组蛋白在体外可以抑制脾细胞增殖和细胞因子IL-2、IFN-γ的分泌。研究结果显示Ha-11在蜱吸血过程中可能具有抗炎作用。
본연구대아주리안비다태Ha-11적편마기인진행료극륭、표체화초보적공능분석。해기인개방열독광（open reading frame, ORF）위372 bp，편마123개안기산，기중함23개안기산적신호태。장거제신호태적편마구기인아극륭지pGEX-4T-1재체，대장간균중유도표체，획득료37 kDa중조단백。항중조Ha-11단백적면역혈청식별반포혈자비적타액선중약11 kDa적천연단백。Real-time PCR결과표명해기인재아주리안비적각발육계단、각조직중균유표체，단이약비표체량최고，이조직중이림파액표체량최고。중조단백재체외가이억제비세포증식화세포인자IL-2、IFN-γ적분비。연구결과현시Ha-11재비흡혈과정중가능구유항염작용。
In the present study, we reported cloning, expression and primary biological identification of Ha-11gene from Hyalomma asiaticum. The ORF of Ha-11gene was 372 bp encoding 123 amino acid residues including a signal peptide of 23 amino acid residues. A 37 kDa recombinant protein was obtained from E.coli DH5αusing pGEX-4T-1 as expression. A specific band about 11kDa was detected in the salivary gland of female ticks in Western blot using anti-recombinant Ha-11serum. The result of Real-time PCR indicated that Ha-11 was expressed in all of developmental stages and tissues of adult female ticks with the highest expression in nymphs and hemolymphs. Recombinant Ha-11 protein significantly suppressed the proliferation of spleen cells and secretion of cytokines including IFN-γand IL-2 invivo. The results obtained here suggested that recombinant protein might be involved in suppression of inflammation.