当前位置: 首页 > 文章 > 鸡IL-18基因的克隆表达及对新城疫疫苗诱导的HI抗体变化的研究 东北农业大学学报 2009,40 (5) 81-85
Position: Home > Articles > Cloning and expression of chicken IL-18 and study on kinetic changes of HI antibody induced by newcastle disease vaccine Journal of Northeast Agricultural University 2009,40 (5) 81-85

鸡IL-18基因的克隆表达及对新城疫疫苗诱导的HI抗体变化的研究

作  者:
姜文博;孙进华;单安山
单  位:
东北农业大学动物营养所
关键词:
鸡白细胞介素18;克隆;表达;新城疫疫苗
摘  要:
通过RT-PCR方法从新城疫Ⅰ系疫苗诱导的11日龄鸡胚脾脏细胞中扩增出鸡白细胞介素18(Chicken interleukin-18,ChIL-18)成熟蛋白基因的cDNA,并将其克隆到pMD18-T载体上。DNA序列测定表明,克隆得到ChIL-18全基因,其编码区大小为597bp,编码198个氨基酸多肽。将该基因片段克隆到原核表达载体pProEXTMHTb构建重组质粒pProEXTMHTb-ChIL-18转化大肠杆菌DH5α中,并用IPTG诱导。重组菌菌体裂解物SDS-PAGE可检测到相对分子量为28.6ku的重组蛋白。观察ChIL-18蛋白对新城疫疫苗诱导的HI抗体动态变化。结果表明,同时接种ChIL-18蛋白和活疫苗的免疫鸡在接种后,所产生的HI抗体效价高于PBS和活疫苗免疫组,并且,HI抗体效价在15d及热应激24h时表现差异显著(P<0.05)。表明ChIL-18蛋白对新城疫疫苗诱导的HI抗体有增强作用。
译  名:
Cloning and expression of chicken IL-18 and study on kinetic changes of HI antibody induced by newcastle disease vaccine
作  者:
JIANG Wenbo, SUN Jinhua, SHAN Anshan (Institute of Animal Nutrition, Northeast Agricultural University, Harbin 150030, China)
关键词:
chicken interleukin-18; clone; expression; New casltle disease virues
摘  要:
Chicken interleukin-18(ChIL-18) mature protein gene was amplified from spleen cells of chicken embryo which were induced by New casltle disease virues by RT-PCR. PCR product was cloned into pMD18-T vector. Sequencing analysis showed that the nucleotide sequence of this ChIL-18 mature protein gene was 597 bp, pProEXTMHTb-ChIL-18 was obtained by subcloning the encodeing region of the ChIL-18 mature protein gene into pProEXTMHTb. The recombinant ChIL-18 (rChIL-18) was expressed efficiently in pProEXTMHTb-ChIL-18 transformed DH5α induced by IPTG. SDS-PAGE analysis showed that the recombinant mature protein had a relative molecular mass of approx in ately 28.6 ku. The effects of the ChIL-18 protein on Newcastle disease vaccine were observed by the kinetic changes of HI antibody. The results showed that ChIL-18 protein could enhance the valence of HI antibody.

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