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表达鸡白细胞介素18重组禽痘病毒的构建及其表达产物生物学活性的检测

DOI: 10.7668/hbnxb.2008.06.002, PP. 6-10

Keywords: 鸡IL-18基因,重组禽痘病毒,构建,生物学活性

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Abstract:

为构建表达鸡IL-18的重组禽痘病毒,将含痘病毒启动子LP2EP2驱动的鸡IL-18基因插入到禽痘病毒转移载体pSY681中,获得重组转移载体pSY681/ChIL-18。将pSY681/ChIL-18转染已感染亲本禽痘病毒S-FPV-017株的鸡胚成纤维细胞,使其在鸡胚成纤维细胞内与禽痘病毒基因组发生同源重组,产生表达鸡IL-18的重组禽痘病毒rFPV-ChIL-18。在含有X-gal的营养琼脂培养基上进行蓝斑筛选后,对重组病毒rFPV-ChIL-18又进行了多次蚀斑克隆。以重组禽痘病毒DNA为模板,利用鸡IL-18基因特异引物进行PCR,扩增出1条约0.6kb的带。收集含鸡IL-18蛋白的细胞上清液进行MTT试验,表达的产物能明显提高鸡淋巴细胞的转化率。

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